U.S. patent number 9,884,861 [Application Number 15/093,354] was granted by the patent office on 2018-02-06 for compositions useful for treating disorders related to kit.
This patent grant is currently assigned to BLUEPRINT MEDICINES CORPORATION. The grantee listed for this patent is BLUEPRINT MEDICINES CORPORATION. Invention is credited to Brian L. Hodous, Joseph L. Kim, Chandrasekhar V. Miduturu, Douglas Wilson, Yulian Zhang.
United States Patent |
9,884,861 |
Hodous , et al. |
February 6, 2018 |
Compositions useful for treating disorders related to kit
Abstract
Compounds and compositions useful for treating disorders related
to KIT and PDFGR are described herein.
Inventors: |
Hodous; Brian L. (Cambridge,
MA), Kim; Joseph L. (Wayland, MA), Miduturu;
Chandrasekhar V. (Cambridge, MA), Wilson; Douglas (Ayer,
MA), Zhang; Yulian (Acton, MA) |
Applicant: |
Name |
City |
State |
Country |
Type |
BLUEPRINT MEDICINES CORPORATION |
Cambridge |
MA |
US |
|
|
Assignee: |
BLUEPRINT MEDICINES CORPORATION
(Cambridge, MA)
|
Family
ID: |
51869029 |
Appl.
No.: |
15/093,354 |
Filed: |
April 7, 2016 |
Prior Publication Data
|
|
|
|
Document
Identifier |
Publication Date |
|
US 20170057953 A1 |
Mar 2, 2017 |
|
Related U.S. Patent Documents
|
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
14517480 |
Oct 17, 2014 |
9334263 |
|
|
|
61892077 |
Oct 17, 2013 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
45/06 (20130101); A61P 17/00 (20180101); C07D
403/14 (20130101); C07D 251/18 (20130101); C07D
417/14 (20130101); A61K 31/553 (20130101); C07D
403/12 (20130101); A61K 31/53 (20130101); A61P
35/02 (20180101); A61K 31/675 (20130101); C07D
471/04 (20130101); A61K 31/635 (20130101); C07F
9/6521 (20130101); A61P 35/00 (20180101); A61K
31/5377 (20130101); A61K 31/5415 (20130101); A61K
31/55 (20130101); C07D 405/14 (20130101); C07D
401/14 (20130101); C07D 413/14 (20130101); C07F
9/65583 (20130101) |
Current International
Class: |
C07D
401/14 (20060101); A61K 31/5377 (20060101); A61K
31/53 (20060101); C07D 471/04 (20060101); C07D
417/14 (20060101); C07D 413/14 (20060101); C07D
403/14 (20060101); C07F 9/6558 (20060101); C07D
251/18 (20060101); C07F 9/6521 (20060101); C07D
405/14 (20060101); C07D 403/12 (20060101); A61K
31/5415 (20060101); A61K 31/553 (20060101); A61K
31/55 (20060101); A61P 35/00 (20060101); A61K
31/675 (20060101); A61K 31/635 (20060101); A61K
45/06 (20060101) |
Field of
Search: |
;544/209,113,182
;540/490,500 ;514/211.05,212.07,231.5,242,245 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
|
|
|
|
|
|
|
0071129 |
|
Nov 2000 |
|
WO |
|
01/25220 |
|
Apr 2001 |
|
WO |
|
2005117909 |
|
Dec 2005 |
|
WO |
|
2007085188 |
|
Aug 2007 |
|
WO |
|
2008005956 |
|
Jan 2008 |
|
WO |
|
2009015254 |
|
Jan 2009 |
|
WO |
|
2009117157 |
|
Sep 2009 |
|
WO |
|
2010022055 |
|
Feb 2010 |
|
WO |
|
2010144345 |
|
Dec 2010 |
|
WO |
|
2011005119 |
|
Jan 2011 |
|
WO |
|
2011103196 |
|
Aug 2011 |
|
WO |
|
2012027495 |
|
Mar 2012 |
|
WO |
|
2014160521 |
|
Oct 2014 |
|
WO |
|
2015057873 |
|
Apr 2015 |
|
WO |
|
2015058129 |
|
Apr 2015 |
|
WO |
|
2016022569 |
|
Feb 2016 |
|
WO |
|
Other References
International Search Report and Written Opinion for International
Application No. PCT/US2014/060746 dated Dec. 17, 2014. cited by
applicant .
International Search Report and Written Opinion for International
Application No. PCT/US2014/061211 dated Oct. 12, 2014. cited by
applicant .
International Search Report and Written Opinion for International
Application No. PCT/US2014/027008 dated Jul. 17, 2014. cited by
applicant .
Quintela et al, "A Ready One-pot Preparation for
Pyrrolo[2,1-f]-[1,2,4]triazine and
Pyrazolo[5,1-c]pyrimido[4,5-e]-[1,2,4]triazine Derivatives"
Tetrahedron (1996) vol. 52, No. 8, pp. 3037-3048. cited by
applicant .
Antonescu, "What lessons can be learned from the GIST paradigm that
can be applied to other kinase-driven cancers" J. Pathol. (2011)
vol. 223, No. 2, pp. 251-261. cited by applicant .
Lee et al. "Correlation of Imatinib Resistance with the Mutational
Status of KIT and ODGFRA Genes in Gastrointestinal Stromal Tumors:
a Meta-analysis" J. Gastrointestin Liver Dis. (2013) vol. 22, No.
4, pp. 413-418. cited by applicant .
Cecil Textbook of Medicine, Edited by Bennet and Plum (1996) 20th
edition, vol. 1, pp. 1004-1010. cited by applicant .
Cohen et al., "The development and therapeutic potential of protein
kinase inhibitors" Current Opinion in Chemical Biology (1999) vol.
3, pp. 459-465. cited by applicant .
Fresheny et al., "Culture of Animal Cells, A Manual of Basic
Technique" Alan R. Liss, Inc. (1983) pp. 1-6. cited by applicant
.
Schnittger et al. "KIT-D816 mutations in AML1-ETO-positive AML are
associated with impaired event-free and overal survival" Blood
(2006) vol. 107, pp. 1791-1799. cited by applicant .
Paschka et al. "Adverse Prognostic Significance of KIT Mutations in
Adult Acute Myeloid Leukemia with inv(16) and t (8;21):A Cancer and
Leukemia Group Study" Journal of Clinical Oncology (2006) vol. 24,
No. 24, pp. 3904-3911. cited by applicant .
Cairoli et al. "Prognostic impact of c-KIT mutations in core
binding factor leukemias: an Italian retrospective study" Blood
(2006) vol. 107, pp. 3463-3468. cited by applicant .
International Search Report for International Application No.
PCT/US2015/043624 dated Oct. 6, 2015. cited by applicant .
Dermer "Another Anniversary for the War on Cancer" Bio/Technology
(1994) vol. 12, pp. 320. cited by applicant.
|
Primary Examiner: Balasubramanian; Venkataraman
Attorney, Agent or Firm: Finnegan, Henderson, Farabow,
Garrett & Dunner, LLP
Parent Case Text
CLAIM OF PRIORITY
This application is a continuation of U.S. Ser. No. 14/517,480
filed Oct. 17, 2014, which claims priority to U.S. Ser. No.
61/892,077 filed Oct. 17, 2013, each of which is incorporated
herein in its entirety.
Claims
We claim:
1. A compound or a pharmaceutically acceptable salt thereof
selected from: ##STR00389## ##STR00390## ##STR00391## ##STR00392##
##STR00393## ##STR00394## ##STR00395## ##STR00396## ##STR00397##
##STR00398## ##STR00399## ##STR00400## ##STR00401## ##STR00402##
##STR00403## ##STR00404## ##STR00405## ##STR00406## ##STR00407##
##STR00408## ##STR00409## ##STR00410## ##STR00411## ##STR00412##
##STR00413## ##STR00414## ##STR00415## ##STR00416## ##STR00417##
##STR00418## ##STR00419## ##STR00420## ##STR00421## ##STR00422##
##STR00423## ##STR00424## ##STR00425## ##STR00426## ##STR00427##
##STR00428## ##STR00429## ##STR00430## ##STR00431## ##STR00432##
##STR00433## ##STR00434## ##STR00435## ##STR00436## ##STR00437##
##STR00438## ##STR00439## ##STR00440## ##STR00441## ##STR00442##
##STR00443## ##STR00444## ##STR00445## ##STR00446## ##STR00447##
##STR00448## ##STR00449## ##STR00450## ##STR00451## ##STR00452##
##STR00453## ##STR00454## ##STR00455## ##STR00456## ##STR00457##
##STR00458## ##STR00459## ##STR00460## ##STR00461##
2. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a compound of claim 1 or a pharmaceutically
acceptable salt thereof.
3. A method of treating mastocytosis comprising administering to a
patient in need thereof a therapeutically effective amount of a
compound of claim 1 or a pharmaceutically acceptable salt thereof
or a pharmaceutical composition of claim 2.
4. The method of claim 3, wherein the mastocytosis is selected from
cutaneous mastocytosis (CM) and systemic mastocytosis (SM).
5. The method of claim 3, wherein the systemic mastocytosis is
selected from indolent systemic mastocytosis (ISM), smoldering
systemic mastocytosis (SSM), aggressive systemic mastocytosis
(ASM), SM with associated hematologic non-mast cell lineage disease
(SM-AHNMD), and mast cell leukemia (MCL).
6. A method of treating gastrointestinal stromal tumor, the method
comprising administering to a patient in need thereof a
therapeutically effective amount of a compound of claim 1 or a
pharmaceutically acceptable salt thereof or a pharmaceutical
composition of claim 2.
7. A method of treating acute myeloid leukemia, the method
comprising administering to a patient in need thereof a
therapeutically effective amount of a compound of claim 1 or a
pharmaceutically acceptable salt thereof or a pharmaceutical
composition of claim 2.
Description
BACKGROUND
The invention relates to compounds and compositions useful for
treating disorders related to KIT and PDGFR.
The enzyme KIT (also called CD117) is a receptor tyrosine kinase
expressed on a wide variety of cell types. The KIT molecule
contains a long extracellular domain, a transmembrane segment, and
an intracellular portion. The ligand for KIT is stem cell factor
(SCF), whose binding to the extracellular domain of KIT induces
receptor dimerization and activation of downstream signaling
pathways. KIT mutations generally occur in the DNA encoding the
juxtumembrane domain (exon 11). They also occur, with less
frequency, in exons 7, 8, 9, 13, 14, 17, and 18. Mutations make KIT
function independent of activation by SCF, leading to a high cell
division rate and possibly genomic instability. Mutant KIT has been
implicated in the pathogenesis of several disorders and conditions
including systemic mastocytosis, GIST (gastrointestinal stromal
tumors), AML (acute myeloid leukemia), melanoma, and seminoma. As
such, there is a need for therapeutic agents that inhibit KIT, and
especially agents that inhibit mutant KIT.
Platelet-derived growth factor receptors (PDGF-R) are cell surface
tyrosine kinase receptors for members of the platelet-derived
growth factor (PDGF) family. PDGF subunits-A and -B are important
factors regulating cell proliferation, cellular differentiation,
cell growth, development and many diseases including cancer. A
PDGFRA D842V mutation has been found in a distinct subset of GIST,
typically from the stomach. The D842V mutation is known to be
associated with tyrosine kinase inhibitor resistance. As such,
there is a need for agents that target this mutation.
SUMMARY OF THE INVENTION
The present invention provides compounds and compositions for
treating or preventing conditions such as mastocytosis by
modulating the activity of Kit, such compounds having the
structural Formula I:
##STR00001## or a pharmaceutically acceptable salt thereof,
wherein:
W is selected from hydrogen, halo and
##STR00002## wherein Ring A is selected from monocyclic or bicyclic
aryl, monocyclic or bicyclic heteroaryl, cycloalkyl and
heterocyclyl;
each X and Y is independently selected from CR.sup.1 and N;
Z is selected from C.sub.1-C.sub.6 alkyl, cycloalkyl, monocyclic or
bicyclic aryl, monocyclic or bicyclic aralkyl, monocyclic or
bicyclic heteroaryl, monocyclic or bicyclic heterocyclyl, and
monocyclic or bicyclic heterocyclylalkyl; wherein each of
C.sub.1-C.sub.6 alkyl, cycloalkyl, monocyclic or bicyclic aryl,
monocyclic or bicyclic aralkyl, monocyclic or bicyclic heteroaryl,
monocyclic or bicyclic heterocyclyl, and monocyclic or bicyclic
heterocyclylalkyl is substituted with 0-5 occurrences of
R.sup.C;
L is selected from a bond, --(C(R.sup.2)(R.sup.2)).sub.m--,
--(C.sub.2-C.sub.6 alkynylene)-, --(C.sub.2-C.sub.6 alkenylene)-,
--(C.sub.1-C.sub.6 haloalkylene)-, --(C.sub.1-C.sub.6
heteroalkylene)-, --(C.sub.1-C.sub.6 hydroxyalkylene)-, --C(O)--,
--O--, --S--, --S(O), --SO.sub.2--, --N(R.sup.2)--,
--O--(C.sub.1-C.sub.6 alkylene)-, --(C.sub.1-C.sub.6 alkylene)-O--,
--N(R.sup.2)--CO--, --CO--N(R.sup.2)--, --(C.sub.1-C.sub.6
alkylene)-N(R.sup.2)--, --N(R.sup.2)--(C.sub.1-C.sub.6 alkylene)-,
--N(R.sup.2)--CO--(C.sub.1-C.sub.6 alkylene)-,
--CO--N(R.sup.2)--(C.sub.1-C.sub.6 alkylene)-,
--N(R.sup.2)--SO.sub.2--, --SO.sub.2--N(R.sup.2)--,
--N(R.sup.2)--SO.sub.2--(C.sub.1-C.sub.6 alkylene)-, and
--SO.sub.2--N(R.sup.2)--(C.sub.1-C.sub.6 alkylene)-;
each R.sup.A and R.sup.B is independently selected from
C.sub.1-C.sub.6 alkyl, halo, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 hydroxyalkyl, C.sub.1-C.sub.6 heteroalkyl,
monocyclic or bicyclic aralkyl, --N(R.sup.2)(R.sup.2), cyano, and
--OR.sup.2;
each R.sup.C is independently selected from C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkynyl, halo, C.sub.1-C.sub.6 heteroalkyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 haloalkoxy,
C.sub.1-C.sub.6 hydroxyalkyl, cycloalkyl, monocyclic or bicyclic
aryl, monocyclic or bicyclic aryloxy, monocyclic or bicyclic
aralkyl, monocyclic or bicyclic heterocyclyl, monocyclic or
bicyclic heterocyclylalkyl, nitro, cyano, --C(O)R.sup.2,
--OC(O)R.sup.2, --C(O)OR.sup.2, --SR.sup.2, --S(O).sub.2R.sup.2,
--S(O).sub.2--N(R.sup.2)(R.sup.2), --(C.sub.1-C.sub.6
alkylene)-S(O).sub.2--N(R.sup.2)(R.sup.2), --N(R.sup.2)(R.sup.2),
--C(O)--N(R.sup.2)(R.sup.2), --N(R.sup.2)(R.sup.2)--C(O)R.sup.2,
--(C.sub.1-C.sub.6 alkylene)-N(R.sup.2)--C(O)R.sup.2,
--NR.sup.2S(O).sub.2R.sup.2, --P(O)(R.sup.2)(R.sup.2), and
--OR.sup.2; wherein each of heteroalkyl, haloalkyl, haloalkoxy,
alkyl, alkynyl, cycloalkyl, aryl, aryloxy, aralkyl, heterocyclyl,
and heterocyclylalkyl is substituted with 0-5 occurrences of
R.sup.a; or 2 R.sup.C together with the carbon atom(s) to which
they are attached form a cycloalkyl or heterocyclyl ring
substituted with 0-5 occurrences of R.sup.a;
each R.sup.1 is independently selected from hydrogen,
C.sub.1-C.sub.6 alkyl, monocyclic aralkyl, C.sub.1-C.sub.6
hydroxyalkyl, halo, C.sub.1-C.sub.6 haloalkyl,
--N(R.sup.2)(R.sup.2), and --OR.sup.2;
each R.sup.2 is independently selected from hydrogen, hydroxyl,
halo, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 hydroxyalkyl, cycloalkyl, cycloalkylalkyl,
heterocyclyl, and heterocyclylalkyl; wherein each of
C.sub.1-C.sub.6 alkyl, cycloalkyl and heterocyclyl is substituted
with 0-5 occurrences of R.sup.b, or 2 R.sup.2 together with the
carbon or nitrogen atom to which they are attached form a
cycloalkyl or heterocyclyl ring;
each R.sup.a and R.sup.b is independently selected from halo,
hydroxyl, --C(O)R', C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.1-C.sub.6
hydroxyalkyl, --NR'R', and cycloalkyl; wherein cycloalkyl is
substituted with 0-5 occurrences of R';
R' is hydrogen, hydroxyl, or C.sub.1-C.sub.6 alkyl; and
m, p, and q are each independently 0, 1, 2, 3, or 4.
Any of the compounds disclosed herein may be used to treat any of
the diseases disclosed herein.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a line graph depicting tumor growth curves for the
different treatment groups: vehicle (), Dasatinib at 25 milligrams
per kilogram (mpk) orally twice a day (po bid) (), Compound 165 at
25 mpk po bid (), Compound 165 at 50 mpk po bid (), and Compound
165 at 100 mpk po bid ().
DETAILED DESCRIPTION OF THE INVENTION
"Aliphatic group" means a straight-chain, branched-chain, or cyclic
hydrocarbon group and includes saturated and unsaturated groups,
such as an alkyl group, an alkenyl group, and an alkynyl group.
"Alkylene" refers to a divalent radical of an alkyl group, e.g.,
--CH.sub.2--, --CH.sub.2CH.sub.2--, and
CH.sub.2CH.sub.2CH.sub.2--.
"Alkenyl" means an aliphatic group containing at least one double
bond.
"Alkoxyl" or "alkoxy" means an alkyl group having an oxygen radical
attached thereto. Representative alkoxyl groups include methoxy,
ethoxy, propyloxy, tert-butoxy and the like.
"Alkyl" refers to a monovalent radical of a saturated straight or
branched hydrocarbon, such as a straight or branched group of 1-12,
1-10, or 1-6 carbon atoms, referred to herein as C.sub.1-C.sub.12
alkyl, C.sub.1-C.sub.10 alkyl, and C.sub.1-C.sub.6 alkyl,
respectively. Exemplary alkyl groups include, but are not limited
to, methyl, ethyl, propyl, isopropyl, 2-methyl-1-propyl,
2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl,
2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl,
3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl,
3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl,
3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, t-butyl,
pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, etc.
"Alkynyl" refers to a straight or branched hydrocarbon chain
containing 2-12 carbon atoms and characterized in having one or
more triple bonds. Examples of alkynyl groups include, but are not
limited to, ethynyl, propargyl, and 3-hexynyl. One of the triple
bond carbons may optionally be the point of attachment of the
alkynyl substituent.
"Hydroxyalkylene" or "hydroxyalkyl" refers to an alkylene or alkyl
moiety in which an alkylene or alkyl hydrogen atom is replaced by a
hydroxyl group. Hydroxyalkylene or hydroxyalkyl includes groups in
which more than one hydrogen atom has been replaced by a hydroxyl
group.
"Aromatic ring system" is art-recognized and refers to a
monocyclic, bicyclic or polycyclic hydrocarbon ring system, wherein
at least one ring is aromatic.
"Aryl" refers to a monovalent radical of an aromatic ring system.
Representative aryl groups include fully aromatic ring systems,
such as phenyl, naphthyl, and anthracenyl, and ring systems where
an aromatic carbon ring is fused to one or more non-aromatic carbon
rings, such as indanyl, phthalimidyl, naphthimidyl, or
tetrahydronaphthyl, and the like.
"Arylalkyl" or "aralkyl" refers to an alkyl moiety in which an
alkyl hydrogen atom is replaced by an aryl group. Aralkyl includes
groups in which more than one hydrogen atom has been replaced by an
aryl group. Examples of "arylalkyl" or "aralkyl" include benzyl,
2-phenylethyl, 3-phenylpropyl, 9-fluorenyl, benzhydryl, and trityl
groups.
"Aryloxy" refers to --O-(aryl), wherein the heteroaryl moiety is as
defined herein.
"Halo" refers to a radical of any halogen, e.g., --F, --Cl, --Br,
or --I.
"Haloalkyl" and "haloalkoxy" refers to alkyl and alkoxy structures
that are substituted with one or more halo groups or with
combinations thereof. For example, the terms "fluoroalkyl" and
"fluoroalkoxy" include haloalkyl and haloalkoxy groups,
respectively, in which the halo is fluorine.
"Heteroalkyl" refers to an optionally substituted alkyl, which has
one or more skeletal chain atoms selected from an atom other than
carbon, e.g., oxygen, nitrogen, sulfur, phosphorus or combinations
thereof. A numerical range may be given, e.g. C.sub.1-C.sub.6
heteroalkyl which refers to the number of carbons in the chain,
which in this example includes 1 to 6 carbon atoms. For example, a
--CH.sub.2OCH.sub.2CH.sub.3 radical is referred to as a "C.sub.3"
heteroalkyl. Connection to the rest of the molecule may be through
either a heteroatom or a carbon in the heteroalkyl chain.
"Carbocyclic ring system" refers to a monocyclic, bicyclic or
polycyclic hydrocarbon ring system, wherein each ring is either
completely saturated or contains one or more units of unsaturation,
but where no ring is aromatic.
"Carbocyclyl" refers to a monovalent radical of a carbocyclic ring
system. Representative carbocyclyl groups include cycloalkyl groups
(e.g., cyclopentyl, cyclobutyl, cyclopentyl, cyclohexyl and the
like), and cycloalkenyl groups (e.g., cyclopentenyl, cyclohexenyl,
cyclopentadienyl, and the like).
"Cycloalkyl" refers to a cyclic, bicyclic, tricyclic, or polycyclic
non-aromatic hydrocarbon groups having 3 to 12 carbons. Any
substitutable ring atom can be substituted (e.g., by one or more
substituents). The cycloalkyl groups can contain fused or spiro
rings. Fused rings are rings that share a common carbon atom.
Examples of cycloalkyl moieties include, but are not limited to,
cyclopropyl, cyclohexyl, methylcyclohexyl, adamantyl, and
norbornyl.
"Cycloalkylalkyl" refers to a -(cycloalkyl)-alkyl radical where
cycloalkyl and alkyl are as disclosed herein. The "cycloalkylalkyl"
is bonded to the parent molecular structure through the cycloalkyl
group.
"Heteroaromatic ring system" is art-recognized and refers to
monocyclic, bicyclic or polycyclic ring system wherein at least one
ring is both aromatic and comprises at least one heteroatom (e.g.,
N, O or S); and wherein no other rings are heterocyclyl (as defined
below). In certain instances, a ring which is aromatic and
comprises a heteroatom contains 1, 2, 3, or 4 ring heteroatoms in
such ring.
"Heteroaryl" refers to a monovalent radical of a heteroaromatic
ring system. Representative heteroaryl groups include ring systems
where (i) each ring comprises a heteroatom and is aromatic, e.g.,
imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, pyrrolyl,
furanyl, thiophenyl pyrazolyl, pyridinyl, pyrazinyl, pyridazinyl,
pyrimidinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl;
(ii) each ring is aromatic or carbocyclyl, at least one aromatic
ring comprises a heteroatom and at least one other ring is a
hydrocarbon ring or e.g., indolyl, isoindolyl, benzothienyl,
benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl,
benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl,
quinazolinyl, quinoxalinyl, carbazolyl, acridinyl, phenazinyl,
phenothiazinyl, phenoxazinyl, pyrido[2,3-b]-1,4-oxazin-3-(4H)-one,
5,6,7,8-tetrahydroquinolinyl and 5,6,7,8-tetrahydroisoquinolinyl;
and (iii) each ring is aromatic or carbocyclyl, and at least one
aromatic ring shares a bridgehead heteroatom with another aromatic
ring, e.g., 4H-quinolizinyl.
"Heterocyclic ring system" refers to monocyclic, bicyclic and
polycyclic ring systems where at least one ring is saturated or
partially unsaturated (but not aromatic) and comprises at least one
heteroatom. A heterocyclic ring system can be attached to its
pendant group at any heteroatom or carbon atom that results in a
stable structure and any of the ring atoms can be optionally
substituted.
"Heterocyclyl" refers to a monovalent radical of a heterocyclic
ring system. Representative heterocyclyls include ring systems in
which (i) every ring is non-aromatic and at least one ring
comprises a heteroatom, e.g., tetrahydrofuranyl, tetrahydropyranyl,
oxetanyl, azetidinyl, tetrahydrothienyl, pyrrolidinyl,
pyrrolidonyl, piperidinyl, pyrrolinyl, decahydroquinolinyl,
oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl,
oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl; (ii) at
least one ring is non-aromatic and comprises a heteroatom and at
least one other ring is an aromatic carbon ring, e.g.,
1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl; and
(iii) at least one ring is non-aromatic and comprises a heteroatom
and at least one other ring is aromatic and comprises a heteroatom,
e.g., 3,4-dihydro-1H-pyrano[4,3-c]pyridine, and
1,2,3,4-tetrahydro-2,6-naphthyridine. In some embodiments,
heterocyclyl can include:
##STR00003##
"Heterocyclylalkyl" refers to an alkyl group substituted with a
heterocycle group.
"Cyano" refers to a --CN radical.
"Nitro" refers to --NO.sub.2.
"Hydroxy" or "hydroxyl" refers to --OH.
"Substituted", whether preceded by the term "optionally" or not,
means that one or more hydrogens of the designated moiety are
replaced with a suitable substituent. Unless otherwise indicated,
an "optionally substituted" group may have a suitable substituent
at each substitutable position of the group, and when more than one
position in any given structure may be substituted with more than
one substituent selected from a specified group, the substituent
may be either the same or different at each position. Combinations
of substituents envisioned under this invention are preferably
those that result in the formation of stable or chemically feasible
compounds. The term "stable", as used herein, refers to compounds
that are not substantially altered when subjected to conditions to
allow for their production, detection, and, in certain embodiments,
their recovery, purification, and use for one or more of the
purposes disclosed herein.
As used herein, the definition of each expression, e.g., alkyl, m,
n, etc., when it occurs more than once in any structure, is
intended to be independent of its definition elsewhere in the same
structure.
Certain compounds of the present invention may exist in particular
geometric or stereoisomeric forms. The present invention
contemplates all such compounds, including cis- and trans-isomers,
R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the
racemic mixtures thereof, and other mixtures thereof, as falling
within the scope of the invention. Additional asymmetric carbon
atoms may be present in a substituent such as an alkyl group. All
such isomers, as well as mixtures thereof, are intended to be
included in this invention.
If, for instance, a particular enantiomer of compound of the
present invention is desired, it may be prepared by asymmetric
synthesis, or by derivation with a chiral auxiliary, where the
resulting diastereomeric mixture is separated and the auxiliary
group cleaved to provide the pure desired enantiomers.
Alternatively, where the molecule contains a basic functional
group, such as amino, or an acidic functional group, such as
carboxyl, diastereomeric salts are formed with an appropriate
optically-active acid or base, followed by resolution of the
diastereomers thus formed by fractional crystallization or
chromatographic means well known in the art, and subsequent
recovery of the pure enantiomers.
Unless otherwise indicated, when a disclosed compound is named or
depicted by a structure without specifying the stereochemistry and
has one or more chiral centers, it is understood to represent all
possible stereoisomers of the compound, as well as enantiomeric
mixtures thereof.
The "enantiomeric excess" or "% enantiomeric excess" of a
composition can be calculated using the equation shown below. In
the example shown below a composition contains 90% of one
enantiomer, e.g., the S enantiomer, and 10% of the other
enantiomer, i.e., the R enantiomer. ee=(90-10)/100=80%.
Thus, a composition containing 90% of one enantiomer and 10% of the
other enantiomer is said to have an enantiomeric excess of 80%.
The compounds or compositions described herein may contain an
enantiomeric excess of at least 50%, 75%, 90%, 95%, or 99% of one
form of the compound, e.g., the S-enantiomer. In other words such
compounds or compositions contain an enantiomeric excess of the S
enantiomer over the R enantiomer.
The compounds described herein may also contain unnatural
proportions of atomic isotopes at one or more of the atoms that
constitute such compounds. For example, the compounds may be
radiolabeled with radioactive isotopes, such as for example
deuterium (.sup.2H), tritium (.sup.3H), carbon-13 (.sup.13C), or
carbon-14 (.sup.14C). All isotopic variations of the compounds
disclosed herein, whether radioactive or not, are intended to be
encompassed within the scope of the present invention. In addition,
all tautomeric forms of the compounds described herein are intended
to be within the scope of the invention.
The compound can be useful as the free base or as a salt.
Representative salts include the hydrobromide, hydrochloride,
sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate,
palmitate, stearate, laurate, benzoate, lactate, phosphate,
tosylate, citrate, maleate, fumarate, succinate, tartrate,
napthylate, mesylate, glucoheptonate, lactobionate, and
laurylsulphonate salts and the like. (See, for example, Berge et
al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19.)
Certain compounds disclosed herein can exist in unsolvated forms as
well as solvated forms, including hydrated forms. The term
"hydrate" or "hydrated" as used herein, refers to a compound formed
by the union of water with the parent compound.
In general, the solvated forms are equivalent to unsolvated forms
and are encompassed within the scope of the present invention.
Certain compounds disclosed herein may exist in multiple
crystalline or amorphous forms. In general, all physical forms are
equivalent for the uses contemplated by the present invention and
are intended to be within the scope of the present invention.
As used herein, the term "patient" refers to organisms to be
treated by the methods of the present invention. Such organisms
preferably include, but are not limited to, mammals (e.g., murines,
simians, equines, bovines, porcines, canines, felines, and the
like), and most preferably includes humans.
As used herein, the term "effective amount" refers to the amount of
a compound (e.g., a compound of the present invention) sufficient
to effect beneficial or desired results. An effective amount can be
administered in one or more administrations, applications or
dosages and is not intended to be limited to a particular
formulation or administration route. As used herein, the term
"treating" includes any effect, e.g., lessening, reducing,
modulating, ameliorating or eliminating, that results in the
improvement of the condition, disease, disorder, and the like, or
ameliorating a symptom thereof.
Compounds
In one embodiment, the invention provides a compound having
structural Formula I:
##STR00004## or a pharmaceutically acceptable salt thereof,
wherein:
W is selected from hydrogen, halo and
##STR00005## wherein Ring A is selected from monocyclic or bicyclic
aryl, monocyclic or bicyclic heteroaryl, cycloalkyl and
heterocyclyl;
each X and Y is independently selected from CR.sup.1 and N;
Z is selected from C.sub.1-C.sub.6 alkyl, cycloalkyl, monocyclic or
bicyclic aryl, monocyclic or bicyclic aralkyl, monocyclic or
bicyclic heteroaryl, monocyclic or bicyclic heterocyclyl, and
monocyclic or bicyclic heterocyclylalkyl; wherein each of
C.sub.1-C.sub.6 alkyl, cycloalkyl, monocyclic or bicyclic aryl,
monocyclic or bicyclic aralkyl, monocyclic or bicyclic heteroaryl,
monocyclic or bicyclic heterocyclyl, and monocyclic or bicyclic
heterocyclylalkyl is substituted with 0-5 occurrences of
R.sup.C;
L is selected from a bond, --(C(R.sup.2)(R.sup.2)).sub.m--,
--(C.sub.2-C.sub.6 alkynylene)-, --(C.sub.2-C.sub.6 alkenylene)-,
--(C.sub.1-C.sub.6 haloalkylene)-, --(C.sub.1-C.sub.6
heteroalkylene)-, --(C.sub.1-C.sub.6 hydroxyalkylene)-, --C(O)--,
--O--, --S--, --S(O), --SO.sub.2--, --N(R.sup.2)--,
--O--(C.sub.1-C.sub.6 alkylene)-, --(C.sub.1-C.sub.6 alkylene)-O--,
--N(R.sup.2)--CO--, --CO--N(R.sup.2)--, --(C.sub.1-C.sub.6
alkylene)-N(R.sup.2)--, --N(R.sup.2)--(C.sub.1-C.sub.6 alkylene)-,
--N(R.sup.2)--CO--(C.sub.1-C.sub.6 alkylene)-,
--CO--N(R.sup.2)--(C.sub.1-C.sub.6 alkylene)-,
--N(R.sup.2)--SO.sub.2--, --SO.sub.2--N(R.sup.2)--,
--N(R.sup.2)--SO.sub.2--(C.sub.1-C.sub.6 alkylene)-, and
--SO.sub.2--N(R.sup.2)--(C.sub.1-C.sub.6 alkylene)-;
each R.sup.A and R.sup.B is independently selected from
C.sub.1-C.sub.6 alkyl, halo, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 hydroxyalkyl, C.sub.1-C.sub.6 heteroalkyl,
monocyclic or bicyclic aralkyl, --N(R.sup.2)(R.sup.2), cyano, and
--OR.sup.2;
each R.sup.C is independently selected from C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkynyl, halo, C.sub.1-C.sub.6 heteroalkyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 haloalkoxy,
C.sub.1-C.sub.6 hydroxyalkyl, cycloalkyl, monocyclic or bicyclic
aryl, monocyclic or bicyclic aryloxy, monocyclic or bicyclic
aralkyl, monocyclic or bicyclic heterocyclyl, monocyclic or
bicyclic heterocyclylalkyl, nitro, cyano, --C(O)R.sup.2,
--OC(O)R.sup.2, --C(O)OR.sup.2, --SR.sup.2, --S(O).sub.2R.sup.2,
--S(O).sub.2--N(R.sup.2)(R.sup.2), --(C.sub.1-C.sub.6
alkylene)-S(O).sub.2--N(R.sup.2)(R.sup.2), --N(R.sup.2)(R.sup.2),
--C(O)--N(R.sup.2)(R.sup.2), --N(R.sup.2)(R.sup.2)--C(O)R.sup.2,
--(C.sub.1-C.sub.6 alkylene)-N(R.sup.2)--C(O)R.sup.2,
--NR.sup.2S(O).sub.2R.sup.2, --P(O)(R.sup.2)(R.sup.2), and
--OR.sup.2; wherein each of heteroalkyl, haloalkyl, haloalkoxy,
alkyl, alkynyl, cycloalkyl, aryl, aryloxy, aralkyl, heterocyclyl,
and heterocyclylalkyl is substituted with 0-5 occurrences of
R.sup.a; or 2 R.sup.C together with the carbon atom(s) to which
they are attached form a cycloalkyl or heterocyclyl ring
substituted with 0-5 occurrences of R.sup.a;
each R.sup.1 is independently selected from hydrogen,
C.sub.1-C.sub.6 alkyl, monocyclic aralkyl, C.sub.1-C.sub.6
hydroxyalkyl, halo, C.sub.1-C.sub.6 haloalkyl,
--N(R.sup.2)(R.sup.2), and --OR.sup.2;
each R.sup.2 is independently selected from hydrogen, hydroxyl,
halo, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 hydroxyalkyl, cycloalkyl, cycloalkylalkyl,
heterocyclyl, and heterocyclylalkyl; wherein each of
C.sub.1-C.sub.6 alkyl, cycloalkyl and heterocyclyl is substituted
with 0-5 occurrences of R.sup.b, or 2 R.sup.2 together with the
carbon or nitrogen atom to which they are attached form a
cycloalkyl or heterocyclyl ring;
each R.sup.a and R.sup.b is independently selected from halo,
hydroxyl, --C(O)R', C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.1-C.sub.6
hydroxyalkyl, --NR'R', and cycloalkyl; wherein cycloalkyl is
substituted with 0-5 occurrences of R';
R' is hydrogen, hydroxyl, or C.sub.1-C.sub.6 alkyl; and
m, p, and q are each independently 0, 1, 2, 3, or 4.
In some embodiments, W is H. In some embodiments, W is halo. In
some embodiments, W is
##STR00006## wherein Ring A is selected from monocyclic or bicyclic
aryl, monocyclic or bicyclic heteroaryl, cycloalkyl and
heterocyclyl. In some embodiments, Ring A is selected from phenyl,
cycloalkyl, monocyclic heteroaryl, and heterocyclyl. In some
embodiments, Ring A is optionally substituted phenyl. In some
embodiments, Ring A is substituted phenyl. In some embodiments,
Ring A is unsubstituted phenyl. In some embodiments, Ring A is
phenyl substituted with halo.
In some embodiments, each R.sup.A is independently selected from
C.sub.1-C.sub.6 alkyl, halo, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 hydroxyalkyl, --N(R.sup.2)(R.sup.2), cyano, and
--OR.sup.2. In some embodiments, R.sup.A is independently selected
from C.sub.1-C.sub.6 alkyl and halo. In some embodiments, q is 0, 1
or 2.
In some embodiments, each R.sup.B is independently selected from
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 hydroxyalkyl,
C.sub.1-C.sub.6 heteroalkyl, --N(R.sup.2)(R.sup.2), cyano and
--OR.sup.2. In some embodiments, each R.sup.B is independently
selected from C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
hydroxyalkyl. In some embodiments, p is 0, 1 or 2. In some
embodiments, p is 0. In some embodiments, p is 1.
In some embodiments, at least one of X and Y is N. In some
embodiments, X and Y are both N. In some embodiments, X and Y are
both CR.sup.1. In some embodiments, X and Y are both CH.
In some embodiments, Z is C.sub.1-C.sub.6 alkyl. In some
embodiments, Z is cycloalkyl, monocyclic or bicyclic aryl,
monocyclic or bicyclic aralkyl, monocyclic or bicyclic heteroaryl,
monocyclic or bicyclic heterocyclyl, or monocyclic or bicyclic
heterocyclylalkyl; wherein each of C.sub.1-C.sub.6 alkyl,
cycloalkyl, monocyclic or bicyclic aryl, monocyclic or bicyclic
aralkyl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic
heterocyclyl, and monocyclic and bicyclic heterocyclylalkyl is
substituted with 0-5 occurrences of R.sup.C. In some embodiments, Z
is bicyclic heteroaryl, bicyclic heterocyclyl, monocyclic
heteroaryl, monocyclic heterocyclylalkyl, or monocyclic aryl. In
some embodiments, Z is aryl or heteroaryl substituted with 0-5
occurrences of R.sup.C. In some embodiments, Z is phenyl
substituted with 0-5 occurrences of R.sup.C. In some embodiments, Z
is phenyl substituted with 1 or 2 occurrences of R.sup.C. In some
embodiments, Z is phenyl substituted with --N(R.sup.2)(R.sup.2),
C.sub.1-C.sub.6 alkyl, or C.sub.1-C.sub.6 hydroxyalkyl. In some
embodiments, Z is heteroaryl substituted with 0-5 occurrences of
R.sup.C. In some embodiments, Z is a heteroaryl ring substituted
with 0-5 occurrences of R.sup.C.
In some embodiments, each R.sup.C is independently selected from
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.1-C.sub.6
hydroxyalkyl, cycloalkyl, monocyclic or bicyclic heterocyclyl,
monocyclic or bicyclic heterocyclylalkyl, --C(O)R.sup.2,
--C(O)OR.sup.2, --SR.sup.2, --S(O).sub.2R.sup.2, and --OR.sup.2;
wherein each of heteroalkyl, alkyl, cycloalkyl, heterocyclyl, and
heterocyclylalkyl is substituted with 0-5 occurrences of
R.sup.a.
In some embodiments, L is selected from a bond, --O--,
--(C(R.sup.2)(R.sup.2)).sub.m--, --O--(C.sub.1-C.sub.6 alkylene)-,
--(C.sub.1-C.sub.6 alkylene)-O--, --(C.sub.2-C.sub.6 alkynylene)-,
--(C.sub.1-C.sub.6 haloalkylene)-, --(C.sub.1-C.sub.6
hydroxyalkylene)-, --S--, --S(O), --SO.sub.2--, and --N(R.sup.2)--.
In some embodiments, L is selected from a bond,
--(C(R.sup.2)(R.sup.2)).sub.m--, --S--, and --SO.sub.2--. In some
embodiments, L is --(C(R.sup.2)(R.sup.2)).sub.m--. In some
embodiments, L is a bond or CH.sub.2. In some embodiments, L is
--(C(R.sup.2)(R.sup.2)).sub.m--, wherein each R.sup.2 is
independently selected from hydrogen, hydroxyl, --NR''R'',
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
hydroxyalkyl, and cycloalkyl; and m is 1 or 2.
In some embodiments, the compound is a compound of Formula II, or a
pharmaceutically acceptable salt thereof, where the substituents
are as defined above.
##STR00007## In some embodiments, L is
--(C(R.sup.2)(R.sup.2)).sub.m--. In some embodiments, X and Y are
CR.sup.1. In some embodiments, Z is phenyl, Z is pyridinyl, Z is
isoxazolyl, Z is pyrazolyl, or Z is dihydroisoquinolinyl.
In some embodiments, Ring A is selected from monocyclic or bicyclic
aryl, monocyclic or bicyclic heteroaryl, cycloalkyl and
heterocyclyl. In some embodiments, Ring A is selected from phenyl,
cycloalkyl, monocyclic heteroaryl, and heterocyclyl. In some
embodiments, Ring A is optionally substituted phenyl. In some
embodiments, Ring A is substituted phenyl. In some embodiments,
Ring A is unsubstituted phenyl. In some embodiments, Ring A is
phenyl substituted with halo.
In some embodiments, each R.sup.A is independently selected from
C.sub.1-C.sub.6 alkyl, halo, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 hydroxyalkyl, --N(R.sup.2)(R.sup.2), cyano, and
--OR.sup.2. In some embodiments, R.sup.A is independently selected
from C.sub.1-C.sub.6 alkyl and halo. In some embodiments, q is 0, 1
or 2.
In some embodiments, each R.sup.B is independently selected from
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 hydroxyalkyl,
C.sub.1-C.sub.6 heteroalkyl, --N(R.sup.2)(R.sup.2), cyano and
--OR.sup.2. In some embodiments, each R.sup.B is independently
selected from C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
hydroxyalkyl. In some embodiments, p is 0, 1 or 2. In some
embodiments, p is 0. In some embodiments, p is 1.
In some embodiments, at least one of X and Y is N. In some
embodiments, X and Y are both N. In some embodiments, X and Y are
both CR.sup.1. In some embodiments, X and Y are both CH.
In some embodiments, Z is C.sub.1-C.sub.6 alkyl. In some
embodiments, Z is cycloalkyl, monocyclic or bicyclic aryl,
monocyclic or bicyclic aralkyl, monocyclic or bicyclic heteroaryl,
monocyclic or bicyclic heterocyclyl, or monocyclic or bicyclic
heterocyclylalkyl; wherein each of C.sub.1-C.sub.6 alkyl,
cycloalkyl, monocyclic or bicyclic aryl, monocyclic or bicyclic
aralkyl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic
heterocyclyl, and monocyclic and bicyclic heterocyclylalkyl is
substituted with 0-5 occurrences of R.sup.C. In some embodiments, Z
is bicyclic heteroaryl, bicyclic heterocyclyl, monocyclic
heteroaryl, monocyclic heterocyclylalkyl, or monocyclic aryl. In
some embodiments, Z is aryl or heteroaryl substituted with 0-5
occurrences of R.sup.C. In some embodiments, Z is phenyl
substituted with 0-5 occurrences of R.sup.C. In some embodiments, Z
is phenyl substituted with 1 or 2 occurrences of R.sup.C. In some
embodiments, Z is phenyl substituted with --N(R.sup.2)(R.sup.2),
C.sub.1-C.sub.6 alkyl, or C.sub.1-C.sub.6 hydroxyalkyl. In some
embodiments, Z is heteroaryl substituted with 0-5 occurrences of
R.sup.C. In some embodiments, Z is a heteroaryl ring substituted
with 0-5 occurrences of R.sup.C.
In some embodiments, each R.sup.C is independently selected from
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.1-C.sub.6
hydroxyalkyl, cycloalkyl, monocyclic or bicyclic heterocyclyl,
monocyclic or bicyclic heterocyclylalkyl, --C(O)R.sup.2,
--C(O)OR.sup.2, --SR.sup.2, --S(O).sub.2R.sup.2, and --OR.sup.2;
wherein each of heteroalkyl, alkyl, cycloalkyl, heterocyclyl, and
heterocyclylalkyl is substituted with 0-5 occurrences of
R.sup.a.
In some embodiments, L is selected from a bond, --O--,
--(C(R.sup.2)(R.sup.2)).sub.m--, --O--(C.sub.1-C.sub.6 alkylene)-,
--(C.sub.1-C.sub.6 alkylene)-O--, --(C.sub.2-C.sub.6 alkynylene)-,
--(C.sub.1-C.sub.6 haloalkylene)-, --(C.sub.1-C.sub.6
hydroxyalkylene)-, --S--, --S(O), --SO.sub.2--, and --N(R.sup.2)--.
In some embodiments, L is selected from a bond,
--(C(R.sup.2)(R.sup.2)).sub.m--, --S--, and --SO.sub.2--. In some
embodiments, L is --(C(R.sup.2)(R.sup.2)).sub.m--. In some
embodiments, L is a bond or CH.sub.2. In some embodiments, L is
--(C(R.sup.2)(R.sup.2)).sub.m--, wherein each R.sup.2 is
independently selected from hydrogen, hydroxyl, --NR''R'',
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
hydroxyalkyl, and cycloalkyl; and m is 1 or 2.
In other embodiments, the compound is a compound of Formula II(a),
or a pharmaceutically acceptable salt thereof, where the
substituents are as defined above.
##STR00008##
In some embodiments, Z is phenyl, Z is pyridinyl, Z is isoxazolyl,
Z is pyrazolyl, or Z is dihydroisoquinolinyl. In some embodiments,
R.sup.C is piperidinyl.
In some embodiments, each R.sup.A is independently selected from
C.sub.1-C.sub.6 alkyl, halo, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 hydroxyalkyl, --N(R.sup.2)(R.sup.2), cyano, and
--OR.sup.2. In some embodiments, R.sup.A is independently selected
from C.sub.1-C.sub.6 alkyl and halo. In some embodiments, q is 0, 1
or 2.
In some embodiments, each R.sup.B is independently selected from
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 hydroxyalkyl,
C.sub.1-C.sub.6 heteroalkyl, --N(R.sup.2)(R.sup.2), cyano and
--OR.sup.2. In some embodiments, each R.sup.B is independently
selected from C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
hydroxyalkyl. In some embodiments, p is 0, 1 or 2. In some
embodiments, p is 0. In some embodiments, p is 1.
In some embodiments, at least one of X and Y is N. In some
embodiments, X and Y are both N. In some embodiments, X and Y are
both CR.sup.1. In some embodiments, X and Y are both CH.
In some embodiments, Z is C.sub.1-C.sub.6 alkyl. In some
embodiments, Z is cycloalkyl, monocyclic or bicyclic aryl,
monocyclic or bicyclic aralkyl, monocyclic or bicyclic heteroaryl,
monocyclic or bicyclic heterocyclyl, or monocyclic or bicyclic
heterocyclylalkyl; wherein each of C.sub.1-C.sub.6 alkyl,
cycloalkyl, monocyclic or bicyclic aryl, monocyclic or bicyclic
aralkyl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic
heterocyclyl, and monocyclic and bicyclic heterocyclylalkyl is
substituted with 0-5 occurrences of R.sup.C. In some embodiments, Z
is bicyclic heteroaryl, bicyclic heterocyclyl, monocyclic
heteroaryl, monocyclic heterocyclylalkyl, or monocyclic aryl. In
some embodiments, Z is aryl or heteroaryl substituted with 0-5
occurrences of R.sup.C. In some embodiments, Z is phenyl
substituted with 0-5 occurrences of R.sup.C. In some embodiments, Z
is phenyl substituted with 1 or 2 occurrences of R.sup.C. In some
embodiments, Z is phenyl substituted with --N(R.sup.2)(R.sup.2),
C.sub.1-C.sub.6 alkyl, or C.sub.1-C.sub.6 hydroxyalkyl. In some
embodiments, Z is heteroaryl substituted with 0-5 occurrences of
R.sup.C. In some embodiments, Z is a heteroaryl ring substituted
with 0-5 occurrences of R.sup.C.
In some embodiments, each R.sup.C is independently selected from
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.1-C.sub.6
hydroxyalkyl, cycloalkyl, monocyclic or bicyclic heterocyclyl,
monocyclic or bicyclic heterocyclylalkyl, --C(O)R.sup.2,
--C(O)OR.sup.2, --SR.sup.2, --S(O).sub.2R.sup.2, and --OR.sup.2;
wherein each of heteroalkyl, alkyl, cycloalkyl, heterocyclyl, and
heterocyclylalkyl is substituted with 0-5 occurrences of
R.sup.a.
The invention also features pharmaceutical compositions comprising
a pharmaceutically acceptable carrier and any compound of Formulas
I-III.
The table below shows the structures of compounds described
herein.
TABLE-US-00001 Compound Number Structure 1 ##STR00009## 2
##STR00010## 3 ##STR00011## 4 ##STR00012## 5 ##STR00013## 6
##STR00014## 7 ##STR00015## 8 ##STR00016## 9 ##STR00017## 10
##STR00018## 11 ##STR00019## 12 ##STR00020## 13 ##STR00021## 14
##STR00022## 15 ##STR00023## 16 ##STR00024## 17 ##STR00025## 18
##STR00026## 19 ##STR00027## 20 ##STR00028## 21 ##STR00029## 22
##STR00030## 23 ##STR00031## 24 ##STR00032## 25 ##STR00033## 26
##STR00034## 27 ##STR00035## 28 ##STR00036## 29 ##STR00037## 30
##STR00038## 31 ##STR00039## 32 ##STR00040## 33 ##STR00041## 34
##STR00042## 35 ##STR00043## 36 ##STR00044## 37 ##STR00045## 38
##STR00046## 39 ##STR00047## 40 ##STR00048## 41 ##STR00049## 42
##STR00050## 43 ##STR00051## 44 ##STR00052## 45 ##STR00053## 46
##STR00054## 47 ##STR00055## 48 ##STR00056## 49 ##STR00057## 50
##STR00058## 51 ##STR00059## 52 ##STR00060## 53 ##STR00061## 54
##STR00062## 55 ##STR00063## 56 ##STR00064## 57 ##STR00065## 58
##STR00066## 59 ##STR00067## 60 ##STR00068## 61 ##STR00069## 62
##STR00070## 63 ##STR00071## 64 ##STR00072## 65 ##STR00073## 66
##STR00074## 67 ##STR00075## 68 ##STR00076## 69 ##STR00077## 70
##STR00078## 71 ##STR00079## 72 ##STR00080## 73 ##STR00081## 74
##STR00082## 75 ##STR00083## 76 ##STR00084## 77 ##STR00085## 78
##STR00086## 79 ##STR00087## 80 ##STR00088## 81 ##STR00089## 82
##STR00090## 83 ##STR00091## 84 ##STR00092## 85 ##STR00093## 86
##STR00094## 87 ##STR00095## 88 ##STR00096## 89 ##STR00097## 90
##STR00098## 91 ##STR00099## 92 ##STR00100## 93 ##STR00101## 94
##STR00102## 95 ##STR00103## 96 ##STR00104## 97 ##STR00105## 98
##STR00106## 99 ##STR00107## 100 ##STR00108## 101 ##STR00109## 102
##STR00110## 103 ##STR00111## 104 ##STR00112## 105 ##STR00113## 106
##STR00114## 107 ##STR00115## 108 ##STR00116## 109 ##STR00117## 110
##STR00118## 111 ##STR00119## 112 ##STR00120## 113 ##STR00121## 114
##STR00122## 115 ##STR00123## 116 ##STR00124## 117 ##STR00125## 118
##STR00126## 119 ##STR00127## 120 ##STR00128## 121 ##STR00129## 122
##STR00130## 123 ##STR00131##
124 ##STR00132## 125 ##STR00133## 126 ##STR00134## 127 ##STR00135##
128 ##STR00136## 129 ##STR00137## 130 ##STR00138## 131 ##STR00139##
132 ##STR00140## 133 ##STR00141## 134 ##STR00142## 135 ##STR00143##
136 ##STR00144## 137 ##STR00145## 138 ##STR00146## 139 ##STR00147##
140 ##STR00148## 141 ##STR00149## 142 ##STR00150## 143 ##STR00151##
144 ##STR00152## 145 ##STR00153## 146 ##STR00154## 147 ##STR00155##
148 ##STR00156## 149 ##STR00157## 150 ##STR00158## 151 ##STR00159##
152 ##STR00160## 153 ##STR00161## 154 ##STR00162## 155 ##STR00163##
156 ##STR00164## 157 ##STR00165## 158 ##STR00166## 159 ##STR00167##
160 ##STR00168## 161 ##STR00169## 162 ##STR00170## 163 ##STR00171##
164 ##STR00172## 165 ##STR00173## 166 ##STR00174## 167 ##STR00175##
168 ##STR00176## 169 ##STR00177## 170 ##STR00178## 171 ##STR00179##
172 ##STR00180## 173 ##STR00181## 174 ##STR00182## 175 ##STR00183##
176 ##STR00184## 177 ##STR00185## 178 ##STR00186## 179 ##STR00187##
180 ##STR00188## 181 ##STR00189## 182 ##STR00190## 183 ##STR00191##
184 ##STR00192## 185 ##STR00193## 186 ##STR00194## 187 ##STR00195##
188 ##STR00196## 189 ##STR00197## 190 ##STR00198## 191 ##STR00199##
192 ##STR00200## 193 ##STR00201## 194 ##STR00202## 195 ##STR00203##
196 ##STR00204## 197 ##STR00205## 198 ##STR00206## 199 ##STR00207##
200 ##STR00208## 201 ##STR00209## 202 ##STR00210## 203 ##STR00211##
204 ##STR00212## 205 ##STR00213## 206 ##STR00214## 207 ##STR00215##
208 ##STR00216## 209 ##STR00217## 210 ##STR00218## 211 ##STR00219##
212 ##STR00220## 213 ##STR00221## 214 ##STR00222## 215 ##STR00223##
216 ##STR00224## 217 ##STR00225## 218 ##STR00226## 219 ##STR00227##
220 ##STR00228## 221 ##STR00229## 222 ##STR00230## 223 ##STR00231##
224 ##STR00232## 225 ##STR00233## 226 ##STR00234## 227 ##STR00235##
228 ##STR00236## 229 ##STR00237## 230 ##STR00238## 231 ##STR00239##
232 ##STR00240## 233 ##STR00241## 234 ##STR00242## 235 ##STR00243##
236 ##STR00244## 237 ##STR00245## 238 ##STR00246## 239 ##STR00247##
240 ##STR00248## 241 ##STR00249## 242 ##STR00250## 243 ##STR00251##
244 ##STR00252## 245 ##STR00253## 246 ##STR00254## 247 ##STR00255##
248 ##STR00256##
249 ##STR00257## 250 ##STR00258## 251 ##STR00259## 252 ##STR00260##
253 ##STR00261## 254 ##STR00262## 255 ##STR00263## 256 ##STR00264##
257 ##STR00265## 258 ##STR00266## 259 ##STR00267## 260 ##STR00268##
261 ##STR00269## 262 ##STR00270## 263 ##STR00271## 264 ##STR00272##
265 ##STR00273## 266 ##STR00274## 267 ##STR00275## 268 ##STR00276##
269 ##STR00277## 270 ##STR00278## 271 ##STR00279## 272 ##STR00280##
273 ##STR00281## 274 ##STR00282## 275 ##STR00283## 276 ##STR00284##
277 ##STR00285##
Synthesis
Compounds of the invention, including salts and N-oxides thereof,
can be prepared using known organic synthesis techniques and can be
synthesized according to any of numerous possible synthetic routes,
such as those in the Schemes below. The reactions for preparing
compounds of the invention can be carried out in suitable solvents
which can be readily selected by one of skill in the art of organic
synthesis. Suitable solvents can be substantially non-reactive with
the starting materials (reactants), the intermediates, or products
at the temperatures at which the reactions are carried out, e.g.,
temperatures which can range from the solvent's freezing
temperature to the solvent's boiling temperature. A given reaction
can be carried out in one solvent or a mixture of more than one
solvent. Depending on the particular reaction step, suitable
solvents for a particular reaction step can be selected by the
skilled artisan.
Preparation of compounds of the invention can involve the
protection and deprotection of various chemical groups. The need
for protection and deprotection, and the selection of appropriate
protecting groups, can be readily determined by one skilled in the
art. The chemistry of protecting groups can be found, for example,
in Wuts and Greene, Protective Groups in Organic Synthesis, 4th
ed., John Wiley & Sons: New Jersey, (2006), which is
incorporated herein by reference in its entirety.
Reactions can be monitored according to any suitable method known
in the art. For example, product formation can be monitored by
spectroscopic means, such as nuclear magnetic resonance (NMR)
spectroscopy (e.g., .sup.1H or .sup.13C), infrared (IR)
spectroscopy, spectrophotometry (e.g., UV-visible), mass
spectrometry (MS), or by chromatographic methods such as high
performance liquid chromatography (HPLC) or thin layer
chromatography (TLC).
Indications
The compounds described herein can be useful for treating
conditions associated with aberrant KIT activity, in humans or
non-humans. Activating mutations in KIT are found in multiple
indications, including systemic mastocytosis, GIST
(gastrointestinal stromal tumors), AML (acute myeloid leukemia),
melanoma, seminoma, intercranial germ cell tumors, and mediastinal
B-cell lymphoma.
Mastocytosis refers to a group of disorders characterized by
excessive mast cell accumulation in one tissue, or in multiple
tissues. Mastocytosis is subdivided into two groups of disorders:
(1) cutaneous mastocytosis (CM) describes forms that are limited to
the skin; and (2) systemic mastocytosis (SM) describes forms in
which mast cells infiltrate extracutaneous organs, with or without
skin involvement. SM is further subdivided into five forms:
indolent (ISM), smoldering (SSM), aggressive (ASM), SM with
associated hemotologic non-mast cell lineage disease (SM-AHNMD),
and mast cell leukemia (MCL).
Diagnosis of systemic mastocytosis is based in part on histological
and cytological studies of bone marrow showing infiltration by mast
cells of frequently atypical morphology, which frequently
abnormally express non-mast cell markers (CD25 and/or CD2).
Diagnosis of SM is confirmed when bone marrow mast cell
infiltration occurs in the context of one of the following: (1)
abnormal mast cell morphology (spindle-shaped cells); (2) elevated
level of serum tryptase above 20 ng/mL; or (3) the presence of the
activating KIT D816V mutation.
Activating mutations at the D816 position are found in the vast
majority of mastocytosis cases (90-98%), with the most common
mutations being D816V and D816H, and D816Y. The D816V mutation is
found in the activation loop of the kinase domain, and leads to
constitutive activation of KIT kinase.
The compounds described herein may also be useful to treat GIST.
Complete surgical resection remains the principal treatment of
choice for patients with a primary GIST. Surgery is effective in
approximately 50% of patients with GIST; of the remaining patients,
tumor recurrence is frequent. Primary treatment with a KIT
inhibitor such as imatinib has also been shown to be sufficient for
initial treatment. However, resistance to imatinib occurs within
months through somatic mutation. These secondary imatinib resistant
mutations are most frequently located on Exon 11, 13, 14, 17 or 18.
Sunitinib is the standard of care second line treatment for most
imatinib resistant tumors and is effective for those containing
mutations in exons 11, 13 and 14. However, secondary KIT mutations
in exons 17 and 18 are resistant to sunitinib treatment and
furthermore, tumors containing tertiary resistance mutations in
exon 17 and 18 emerge several months after sunitinib treatment.
Regorafenib has shown promising results in a phase 3 clinical trial
of imatinib, sunitinib resistant GISTs with activity against
several but not all exon 17 and 18 mutations, of which D816 is one.
Thus, there is a need for therapeutic agents to treat GIST patients
with exon 17 mutations not addressed by regorafenib.
In addition to the use of the compounds described herein as single
agents in the refractory GIST setting, the use of combinations of
imatinib, sunitinib and/or regorafenib with the compounds disclosed
herein may allow for the prevention of emergence of resistance to
exon 17 mutations.
There is a subset of GIST patients with a D842V mutation in
PDGFR.alpha.; this subgroup of GIST patients can be stratified by
identifying this mutation. This subset of patients is refractory to
all tyrosine kinase inhibitors currently available. The compounds
described herein, due to their activity against PDGFR.alpha. D842V,
can be useful in treating these patients.
The compounds described herein may also be useful in treating AML.
AML patients harbor KIT mutations as well, with the majority of
these mutations at the D816 position.
In addition, mutations in KIT have been linked to Ewing's sarcoma,
DLBCL (diffuse large B cell lymphoma), dysgerminoma, MDS
(myelodysplastic syndrome), NKTCL (nasal NK/T-cell lymphoma), CMML
(chronic myelomonocytic leukemia), and brain cancers.
The compounds disclosed herein may be used to treat conditions
associated with the KIT mutations in Exon 9, Exon 11, Exon 13, Exon
14, Exon 17 and/or Exon 18. They may also be used to treat
conditions associated with wild-type KIT. The compounds described
herein may be used as single agents to treat the conditions
described herein, or they may be used in combination with other
therapeutic agents, including, without limitation, imatinib,
sunitinib and regorafenib. Other agents include the compounds
described in WO 2014/039714 and WO 2014/100620.
Compounds described herein can be active against one or more KIT
mutations in Exon 17 (e.g., D816V, D816Y, D816F, D816K, D816H,
D816A, D816G, D820A, D820E, D820G, N822K, N822H, Y823D, and A829P),
and much less active against wild-type KIT. These compounds can be
administered in combination with an agent that is (a) active
against other activating mutations of KIT, such as Exon 9 and 11
mutations, but (b) not active against the Exon 17 mutations. Such
agents include imatinib, sunitinib, and regorafenib. The
combination of the compound and the agent will thus inhibit Exon 17
mutant KIT, as well as inhibiting Exon 9/11 mutant KIT. The
compound and agent can be co-administered, or administered in an
alternating regimen. That is, the Exon 17 mutant KIT inhibitor can
be administered alone for a period of time; then the Exon 9/11
mutant KIT inhibitor can be administered alone for a period of time
following. This cycle may then be repeated. It is believed that
such a regimen could slow the development of resistance to the Exon
17 mutant KIT inhibitor and/or the Exon 9/11 mutant KIT
inhibitor.
In addition, compounds described herein that can be selective for
Exon 17 KIT mutations can be administered with agents that are
active against Exon 9/11 mutations, in combination with a third
agent that covers mutations that are missed with the two-way combo.
The combination of the three agents could inhibit a spectrum of KIT
mutations, as well as wild-type KIT in some instances. The agents
could be administered simultaneously, or in an alternating regimen.
They can be administered one at a time, or two agents can be
administered together for a period of time; then the third agent
can be administered alone for a following period of time. It is
believed that such a regimen could slow the development of
resistance to the mutant KIT inhibitors.
Pharmaceutical Compositions
While it is possible for a compound disclosed herein to be
administered alone, it is preferable to administer the compound as
a pharmaceutical formulation, where the compound is combined with
one or more pharmaceutically acceptable excipients or carriers. The
compounds disclosed herein may be formulated for administration in
any convenient way for use in human or veterinary medicine. In
certain embodiments, the compound included in the pharmaceutical
preparation may be active itself, or may be a prodrug, e.g.,
capable of being converted to an active compound in a physiological
setting.
The phrase "pharmaceutically acceptable" is employed herein to
refer to those compounds, materials, compositions, and/or dosage
forms which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of human beings and
animals without excessive toxicity, irritation, allergic response,
or other problem or complication, commensurate with a reasonable
benefit/risk ratio.
Examples of pharmaceutically acceptable carriers include: (1)
sugars, such as lactose, glucose and sucrose; (2) starches, such as
corn starch and potato starch; (3) cellulose, and its derivatives,
such as sodium carboxymethyl cellulose, ethyl cellulose and
cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin;
(7) talc; (8) excipients, such as cocoa butter and suppository
waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil,
sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such
as propylene glycol; (11) polyols, such as glycerin, sorbitol,
mannitol and polyethylene glycol; (12) esters, such as ethyl oleate
and ethyl laurate; (13) agar; (14) buffering agents, such as
magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16)
pyrogen-free water; (17) isotonic saline; (18) Ringer's solution;
(19) ethyl alcohol; (20) phosphate buffer solutions; (21)
cyclodextrins such as Captisol.RTM.; and (22) other non-toxic
compatible substances employed in pharmaceutical formulations.
Examples of pharmaceutically acceptable antioxidants include: (1)
water soluble antioxidants, such as ascorbic acid, cysteine
hydrochloride, sodium bisulfate, sodium metabisulfite, sodium
sulfite and the like; (2) oil-soluble antioxidants, such as
ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol,
and the like; and (3) metal chelating agents, such as citric acid,
ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid,
phosphoric acid, and the like.
Solid dosage forms (e.g., capsules, tablets, pills, dragees,
powders, granules and the like) can include one or more
pharmaceutically acceptable carriers, such as sodium citrate or
dicalcium phosphate, and/or any of the following: (1) fillers or
extenders, such as starches, lactose, sucrose, glucose, mannitol,
and/or silicic acid; (2) binders, such as, for example,
carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,
sucrose and/or acacia; (3) humectants, such as glycerol; (4)
disintegrating agents, such as agar-agar, calcium carbonate, potato
or tapioca starch, alginic acid, certain silicates, and sodium
carbonate; (5) solution retarding agents, such as paraffin; (6)
absorption accelerators, such as quaternary ammonium compounds; (7)
wetting agents, such as, for example, cetyl alcohol and glycerol
monostearate; (8) absorbents, such as kaolin and bentonite clay;
(9) lubricants, such a talc, calcium stearate, magnesium stearate,
solid polyethylene glycols, sodium lauryl sulfate, and mixtures
thereof; and (10) coloring agents.
Liquid dosage forms can include pharmaceutically acceptable
emulsions, microemulsions, solutions, suspensions, syrups and
elixirs. In addition to the active ingredient, the liquid dosage
forms may contain inert diluents commonly used in the art, such as,
for example, water or other solvents, solubilizing agents and
emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, oils (in particular,
cottonseed, groundnut, corn, germ, olive, castor and sesame oils),
glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty
acid esters of sorbitan, and mixtures thereof.
Suspensions, in addition to the active compounds, may contain
suspending agents as, for example, ethoxylated isostearyl alcohols,
polyoxyethylene sorbitol and sorbitan esters, microcrystalline
cellulose, aluminum metahydroxide, bentonite, agar-agar and
tragacanth, and mixtures thereof.
Ointments, pastes, creams and gels may contain, in addition to an
active compound, excipients, such as animal and vegetable fats,
oils, waxes, paraffins, starch, tragacanth, cellulose derivatives,
polyethylene glycols, silicones, bentonites, silicic acid, talc and
zinc oxide, or mixtures thereof.
Powders and sprays can contain, in addition to an active compound,
excipients such as lactose, talc, silicic acid, aluminum hydroxide,
calcium silicates and polyamide powder, or mixtures of these
substances. Sprays can additionally contain customary propellants,
such as chlorofluorohydrocarbons and volatile unsubstituted
hydrocarbons, such as butane and propane.
The formulations may conveniently be presented in unit dosage form
and may be prepared by any methods well known in the art of
pharmacy. The amount of active ingredient which can be combined
with a carrier material to produce a single dosage form will vary
depending upon the host being treated, the particular mode of
administration. The amount of active ingredient that can be
combined with a carrier material to produce a single dosage form
will generally be that amount of the compound which produces a
therapeutic effect.
Dosage forms for the topical or transdermal administration of a
compound of this invention include powders, sprays, ointments,
pastes, creams, lotions, gels, solutions, patches and inhalants.
The active compound may be mixed under sterile conditions with a
pharmaceutically acceptable carrier, and with any preservatives,
buffers, or propellants that may be required.
When the compounds disclosed herein are administered as
pharmaceuticals, to humans and animals, they can be given per se or
as a pharmaceutical composition containing, for example, 0.1 to
99.5% (more preferably, 0.5 to 90%) of active ingredient in
combination with a pharmaceutically acceptable carrier.
The formulations can be administered topically, orally,
transdermally, rectally, vaginally, parentally, intranasally,
intrapulmonary, intraocularly, intravenously, intramuscularly,
intraarterially, intrathecally, intracapsularly, intradermally,
intraperitoneally, subcutaneously, subcuticularly, or by
inhalation.
Dosages
Actual dosage levels of the active ingredients in the
pharmaceutical compositions of this invention may be varied so as
to obtain an amount of the active ingredient that is effective to
achieve the desired therapeutic response for a particular patient,
composition, and mode of administration, without being toxic to the
patient.
The selected dosage level will depend upon a variety of factors
including the activity of the particular compound disclosed herein
employed, or the ester, salt or amide thereof, the route of
administration, the time of administration, the rate of excretion
of the particular compound being employed, the duration of the
treatment, other drugs, compounds and/or materials used in
combination with the particular compound employed, the age, sex,
weight, condition, general health and prior medical history of the
patient being treated, and like factors well known in the medical
arts.
A physician or veterinarian having ordinary skill in the art can
readily determine and prescribe the effective amount of the
pharmaceutical composition required. For example, the physician or
veterinarian could start doses of the compounds of the invention
employed in the pharmaceutical composition at levels lower than
that required in order to achieve the desired therapeutic effect
and gradually increase the dosage until the desired effect is
achieved.
In general, a suitable daily dose of a compound of the invention
will be that amount of the compound that is the lowest dose
effective to produce a therapeutic effect. Such an effective dose
will generally depend upon the factors described above. Generally,
intravenous, intracerebroventricular and subcutaneous doses of the
compounds of this invention for a patient will range from about
0.0001 to about 100 mg per kilogram of body weight per day. If
desired, the effective daily dose of the active compound may be
administered as two, three, four, five, six or more sub-doses
administered separately at appropriate intervals throughout the
day, optionally, in unit dosage forms. In some embodiments, the
dose for humans will be 100-1000 mg, or 400-800 mg, administered
twice daily; or 400-1000 mg, administered once daily.
Examples
The following examples are intended to be illustrative, and are not
meant in any way to be limiting.
The below Schemes are meant to provide general guidance in
connection with preparing the compounds of the invention. One
skilled in the art would understand that the preparations shown in
the Schemes can be modified or optimized using general knowledge of
organic chemistry to prepare various compounds of the
invention.
##STR00286## ##STR00287##
Scheme 1 schematically depicts synthetic protocol 1. Triazine (B)
can be reacted with amine (A, Z is aryl or heteroaryl) under
nucleophilic aromatic substitution reaction conditions using an
amine base such as diisopropylethylamine (DIPEA) or triethylamine
(TEA) in a polar solvent such as dioxane to provide the
amine-substituted triazine (C). The amine-substituted triazine (C)
can be substituted with piperazine (D, X and Y are --CH--) under
nucleophilic aromatic substitution reaction conditions using an
amine base such as diisopropylethylamine (DIPEA) or triethylamine
(TEA) in a polar solvent such as dioxane to provide the
piperazine-substituted triazine (E). Piperazine-substituted
heteroaryl (E) can be coupled to the organozinc bromide (F) using
Negishi coupling conditions to provide the substituted triazine
(G). As shown below, Compound 186 was prepared using synthetic
protocol 1.
Synthesis of
N-(1-((S)-morpholin-2-ylmethyl)-1H-pyrazol-4-yl)-4-(4-(5-((R)-1-phenyleth-
yl)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-amine (Compound
186)
##STR00288## ##STR00289##
Step 1: Synthesis of (S)-tert-butyl
2-((methylsulfonyloxy)methyl)morpholine-4-carboxylate
##STR00290##
To a solution of (S)-tert-butyl
2-(hydroxymethyl)morpholine-4-carboxylate (3.0 g, 13.8 mmol) and
triethylamine (4.2 g, 41.4 mmol) in dichloromethane (80 mL) at
0.degree. C. was dropwise added mesyl chloride (1.9 g, 16.5 mmol).
The reaction mixture was stirred at 0.degree. C. for 2 hour, and
then diluted with dichloromethane (100 mL). The organic layers were
washed with water (100 mL) and brine (100 mL), dried over sodium
sulfate, filtered and concentrated to afford (S)-tert-butyl
2-((methylsulfonyloxy)methyl)morpholine-4-carboxylate as a purple
oil (4.0 g, 98%), which was directly used in the next step without
further purification. MS (ES+) C.sub.11H.sub.21NO.sub.6S requires:
295. found: 240 [M+H-56].sup.+.
Step 2: Synthesis of (S)-tert-butyl
2-((4-nitro-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
##STR00291##
To a solution of (S)-tert-butyl
2-((methylsulfonyloxy)methyl)morpholine-4-carboxylate (2.8 g, 9.5
mmol) and 4-nitro-1H-pyrazole (715 mg, 6.3 mmol) in acetonitrile
(100 mL) was added cesium carbonate (6.2 g, 19.0 mmol). The
reaction mixture was stirred at 55.degree. C. overnight and then
concentrated. The residue was dissolved in ethyl acetate (100 mL),
washed with water (50 mL), dried over sodium sulfate, filtered and
concentrated. The crude sample was purified by silica gel
chromatography, eluting with petroleum ether:ethyl acetate=8:1, to
afford (S)-tert-butyl
2-((4-nitro-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate as a
yellow oil (2.12 g, 71%). MS (ES+) C.sub.13H.sub.20N.sub.4O.sub.5
requires: 312. found: 257 [M+H-56].sup.+.
Step 3: Synthesis of (S)-tert-butyl
2-((4-amino-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
##STR00292##
To a solution of (S)-tert-butyl
2-((4-nitro-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate (3.2 g,
10.2 mmol) in methanol (100 mL) was added Pd/C (600 mg). The
mixture was stirred under 1 atm H.sub.2 at room temperature
overnight, and then filtrated through a pad of celite. The filtrate
was concentrated to afford (S)-tert-butyl
2-((4-amino-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate. MS
(ES+) C.sub.13H.sub.22N.sub.4O.sub.3 requires: 282. found: 283
[M+H].sup.+.
Step 4: Synthesis of (S)-tert-butyl
2-((4-(4-chloro-1,3,5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholin-
e-4-carboxylate
##STR00293##
To a solution of 2,4-dichloro-1,3,5-triazine (1.8 g, 6.4 mmol) in
dioxane (20 mL) was added diisopropylethylamine (4 mL), followed by
the addition of (S)-tert-butyl
2-((4-amino-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate (1.2 g,
7.2 mmol). The reaction mixture was stirred at 110.degree. C. for 1
hour, and LCMS (liquid chromatography-mass spectrometry) showed the
reaction was completed. The solvents were removed under reduced
pressure, and the residue was purified by silica gel
chromatography, eluting with petroleum ether:ethyl acetate=5:1, to
afford (S)-tert-butyl
2-((4-(4-chloro-1,3,5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholin-
e-4-carboxylate as a yellow solid (1.8 g, 70%). MS (ES+)
C.sub.16H.sub.22ClN.sub.7O.sub.3 requires: 395, 396. found: 396,
397 [M+H].sup.+.
Step 5: Synthesis of 5-bromo-2-(piperazin-1-yl)pyrimidine HCl
Salt
##STR00294##
A solution of tert-butyl
4-(5-bromopyrimidin-2-yl)piperazine-1-carboxylate (5 g, 14.6 mmol)
in 4 M HCl-dioxane (50 mL) was stirred at room temperature for 1
hour. The reaction mixture was concentrated to give
5-bromo-2-(piperazin-1-yl)pyrimidine HCl salt (crude, 3.3 g, 94%)
as a white solid. MS (ES+) C.sub.8H.sub.11BrN.sub.4 requires: 242,
244. found: 243, 245 [M+H].sup.+.
Step 6: Synthesis of (S)-tert-butyl
2-((4-(4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylamin-
o)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
##STR00295##
To a solution of (S)-tert-butyl
2-((4-(4-chloro-1,3,5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholin-
e-4-carboxylate (1.8 g, 4.5 mmol) in dioxane (50 mL) was added
diisopropylethylamine (4 mL) and
5-bromo-2-(piperazin-1-yl)pyrimidine (1.3 g, 5.5 mmol). The
reaction mixture was stirred at room temperature for 2 hours. The
solvents were removed, and the residue was washed with methanol (10
mL) to afford (S)-tert-butyl
2-((4-(4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylamin-
o)-1H-pyrazol-1-yl)methyl)-morpholine-4-carboxylate as a white
solid (2.4 g, 89%), which was directly used in the next step
without further purification. MS (ES+)
C.sub.24H.sub.32BrN.sub.11O.sub.3 requires: 601, 603. found: 602,
604 [M+H].sup.+.
Step 7: Synthesis of (2S)-tert-butyl
2-((4-(4-(4-(5-(1-phenylethyl)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazi-
n-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
##STR00296##
To a solution of (S)-tert-butyl
2-((4-(4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylamin-
o)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate (100 mg, 0.17
mmol) and Pd(Amphos)Cl.sub.2 (11.7 mg, 0.17 mmol) in THF (2 mL,
dried) was dropwise added a solution of (1-phenylethyl)zinc(II)
bromide in THF (7.0 mL, 0.5 M, 3.4 mmol). The reaction mixture was
stirred at 70.degree. C. for 1 hour under N.sub.2, then cooled to
room temperature and diluted with ethyl acetate (50 mL). After
filtration, the filtrate was concentrated to afford (2S)-tert-butyl
2-((4-(4-(4-(5-(1-phenylethyl)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazi-
n-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate as a
white solid (130 mg, crude), which was directly used in the next
step without further purification. MS (ES+)
C.sub.32H.sub.41N.sub.11O.sub.3 requires: 627. found: 628
[M+H].sup.+.
Step 8: Synthesis of
N-(1-((S)-morpholin-2-ylmethyl)-1H-pyrazol-4-yl)-4-(4-(5-((R)-1-phenyleth-
yl)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-amine
##STR00297##
To a solution of (2S)-tert-butyl
2-((4-(4-(4-(5-(1-phenylethyl)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazi-
n-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate (130
mg, crude) in dichloromethane (6 mL) was dropwise added
trifluoroacetic acid (2 mL). The reaction mixture was stirred at
room temperature for 0.5 hour and then concentrated. The residue
was purified by Prep-HPLC to provide
N-(1-((S)-morpholin-2-ylmethyl)-1H-pyrazol-4-yl)-4-(4-(5-(1-phenylethyl)p-
yrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-amine as a white
solid, which was then separated by Chiral-HPLC to afford
N-(1-((S)-morpholin-2-ylmethyl)-1H-pyrazol-4-yl)-4-(4-(5-((R)-1-phenyleth-
yl)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-amine as a white
solid (15.4 mg, 17%).
##STR00298##
Scheme 2 schematically depicts synthetic protocol 2. Triazine (B)
can be reacted with amine (A, Z is aryl or heteroaryl) under
nucleophilic aromatic substitution reaction conditions using an
amine base such as diisopropylethylamine (DIPEA) or triethylamine
(TEA) in a polar solvent such as dioxane to provide the
amine-substituted triazine (C). The amine-substituted triazine (C)
can be substituted with piperazine (D, X and Y are --CH--, A is
aryl, and L is C.sub.1-6 alkyl, cycloalkyl, C.sub.1-6 haloalkyl, or
sulfur) under nucleophilic aromatic substitution reaction
conditions using an amine base such as diisopropylethylamine
(DIPEA) or triethylamine (TEA) in a polar solvent such as dioxane
to provide the piperazine-substituted triazine (E). As shown below,
Compound 165 was prepared using synthetic protocol 2.
Synthesis of
(R)-1-(4-((4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-y-
l)amino)benzyl)piperidin-3-ol (Compound 165)
##STR00299## ##STR00300##
Step 1: Synthesis of (R)-1-(4-nitrophenyl)piperidin-3-ol
##STR00301##
4-Fluoronitrobenze (1.5 g, 10.6 mmole), (R)-piperidin-3-ol HCl
(1.76 g, 12.8 mmole) and potassium carbonate (4.41 g, 31.9 mmole)
were combined in 10 mL DMF and heated to 60.degree. C. overnight.
The reaction mixture was cooled to room temperature and poured into
.about.75 mL water; the suspension was stirred at room temperature
for 30 minutes. The mixture was then filtered, washed with water
and suction dried to give 2.25 g (95%) of
(R)-1-(4-nitrophenyl)piperidin-3-ol as a yellow solid.
Step 2: Synthesis of (R)-1-(4-aminophenyl)piperidin-3-ol
##STR00302##
To a solution of (R)-1-(4-nitrophenyl)piperidin-3-ol was added
Pd/C. The mixture was stirred under 1 atm H.sub.2 at room
temperature overnight, and then filtrated through a pad of Celite.
The filtrate was concentrated to afford
(R)-1-(4-aminophenyl)piperidin-3-ol.
Step 3: Synthesis of
(R)-1-(4-((4-chloro-1,3,5-triazin-2-yl)amino)phenyl)piperidin-3-ol
##STR00303##
To a solution of 2,4-dichloro-1,3,5-triazine in dioxane was added
diisopropylethylamine, followed by the addition of
(R)-1-(4-aminophenyl)piperidin-3-ol. The reaction mixture was
stirred at 110.degree. C. for 1 hour, and LCMS (liquid
chromatography-mass spectrometry) showed the reaction was
completed. The solvents were removed under reduced pressure, and
the residue was purified by silica gel chromatography, eluting with
petroleum ether:ethyl acetate=5:1, to afford
(R)-1-(4-((4-chloro-1,3,5-triazin-2-yl)amino)phenyl)piperidin-3-ol-
.
Step 4: Synthesis of
(R)-1-(4-((4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-y-
l)amino)benzyl)piperidin-3-ol
##STR00304##
To a solution of
(R)-1-(4-((4-chloro-1,3,5-triazin-2-yl)amino)phenyl)piperidin-3-ol
in dioxane was added diisopropylethylamine
5-benzyl-2-(piperazin-1-yl)pyrimidine. The reaction mixture was
stirred at room temperature for 2 hours. The solvents were removed,
and the residue was washed with methanol (10 mL) to afford
(R)-1-(4-((4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-y-
l)amino)benzyl)piperidin-3-ol (Compound 165).
##STR00305##
Scheme 3 schematically depicts synthetic protocol 3. Triazine (B)
can be reacted with piperizine (A, X and Y are --CH--, A is aryl,
and L is C.sub.1-6 alkyl, cycloalkyl, C.sub.1-6haloalkyl, or
sulfur) under nucleophilic aromatic substitution reaction
conditions using an amine base such as diisopropylethylamine
(DIPEA) or triethylamine (TEA) in a polar solvent such as dioxane
to provide the piperizine-substituted triazine (C). The
piperizine-substituted triazine (C) can be substituted with amine
(D, Z is aryl or heteroaryl) under nucleophilic aromatic
substitution reaction conditions using an amine base such as
diisopropylethylamine (DIPEA) or triethylamine (TEA) in a polar
solvent such as dioxane to provide the piperazine-substituted
triazine (E). As shown below, Compound 64 was prepared using
synthetic protocol 3.
Synthesis of
1-(4-(4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylamin-
o)-1H-pyrazol-1-yl)ethane-1,2-diol
##STR00306## ##STR00307## ##STR00308##
Step 1: Synthesis of (2,2-dimethyl-1,3-dioxolan-4-yl)methyl
4-methylbenzenesulfonate
##STR00309##
To a solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methanol (5.28 g,
40.0 mmol), triethyl amine (12.12 g, 120.0 mmol) and
4-dimethylaminopyridine (500 mg, 4.0 mmol) in dichloromethane (200
mL) was added tosyl chloride (15.2 g, 80.0 mmol). The reaction
mixture was stirred at RT overnight, then quenched with water (200
mL) and extracted with ethyl acetate. The organic layers were
separated, combined, washed with water (200 mL) and brine (200 mL),
dried over sodium sulfate, filtered, and concentrated. The residue
was purified by silica gel chromatography, eluting with ethyl
acetate/petroleum ether=1/8, to afford
(2,2-dimethyl-1,3-dioxolan-4-yl)methyl 4-methylbenzenesulfonate
(6.1 g, yield 53%), MS (ES+) C.sub.13H.sub.18O.sub.5S requires:
286. found: 287 [M+H].sup.+.
Step 2: Synthesis of
1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-4-nitro-1H-pyrazole
##STR00310##
A solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methyl
4-methylbenzenesulfonate (1.57 g, 5.5 mmol), 4-nitro-1H-pyrazole
(0.57 g, 5.0 mmol) and cesium carbonate (4.89 g, 15.0 mmol) in
acetonitrile (30 mL) was stirred at 70.degree. C. under N.sub.2
overnight. The reaction mixture was filtered, and the resulting
filtrate was concentrated. The residue was diluted with ethyl
acetate (50 mL), washed with water (50 mL.times.2) and brine (50
mL), dried over sodium sulfate, filtered, and concentrated to
afford
1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-4-nitro-1H-pyrazole
(1.01 g, crude), which was directly used in the next step without
further purification. MS (ES+) C.sub.9H.sub.13N.sub.3O.sub.4
requires: 227. found: 228 [M+H].sup.+.
Step 3: Synthesis of
1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-pyrazol-4-amine
##STR00311##
A mixture of
1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-4-nitro-1H-pyrazole (2.0
g, 9.5 mmol) and Pd/C (400 mg) in methanol (40 mL) was stirred at
RT under H.sub.2 overnight. LCMS showed the reaction was completed.
The reaction mixture was filtered through a pad of celite, and the
filtrate was concentrated to give
1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-pyrazol-4-amine (1.7
g, crude) as a purple oil, which was directly used in the next step
without further purification. MS (ES+)
C.sub.9H.sub.15N.sub.3O.sub.2 requires: 197. found: 198
[M+H].sup.+.
Step 4: Synthesis of tert-butyl 4-(5-bromopyrimidin-2-yl)
piperazine-1-carboxylate
##STR00312##
To a solution of 5-bromo-2-chloropyrimidine (50.0 g, 258 mmol) and
1-tert-butoxycarbonylpiperazine (72.2 g, 387 mmol) in 1,4-dioxane
(500 mL) was added potassium carbonate (67.8 g, 491 mmol), and the
mixture was stirred under reflux for 1.5 h. The mixture was diluted
with water (500 mL) and extracted with diethyl ether (1000 mL*2).
The combined organic layers were dried over sodium sulfate,
filtered and concentrated. The residue was purified with silica gel
chromatography (elute:hexane:ethyl acetate=8:1 to 4:1) to give
tert-butyl 4-(5-bromopyrimidin-2-yl) piperazine-1-carboxylate (70.5
g, 80%) as a white solid. MS (ES+) C.sub.13H.sub.19BrN.sub.4O.sub.2
requires: 342. found: 243 [M+H-100].sup.+.
Step 5: Synthesis of Benzylzinc(II) Bromide
##STR00313##
To a suspension mixture of zinc powder (active, 65 g, 1.0 mol) in
dry THF (200 mL) was dropwise added 1,2-dibromoethane (3.5 mL, 40
mmol) at 65.degree. C. under nitrogen atmosphere, followed by the
addition of chlorotrimethylsilane (87 mg, 80 mmol). The mixture was
then stirred at 60.degree. C. for another 1 h. Subsequently,
(bromomethyl)benzene (60 mL, 500 mmol) was dropwise added, and the
suspension was stirred at 60.degree. C. for another 1 h. The
reaction mixture was directly used in the next step.
Step 6: Synthesis of tert-butyl
4-(5-benzylpyrimidin-2-yl)piperazine-1-carboxylate
##STR00314##
To a solution of tert-butyl
4-(5-bromopyrimidin-2-yl)piperazine-1-carboxylate (12.0 g, 35.0
mmol) and tetrakis(triphenylphosphine)palladium (2.0 g, 1.7 mmol)
in THF (200 mL, dry) was added dropwise a solution of
benzylzinc(II) bromide in THF (140 mL, 0.5 M, 70 mmol). The
reaction mixture was stirred at 70.degree. C. for 4 h under
N.sub.2, cooled to RT, then diluted with ethyl acetate (300 mL),
filtered and concentrated. The residue was purified by silica gel
chromatography, eluting with ethyl acetate/petroleum
ether=1/20.about.1/10, to give -butyl
4-(5-benzylpyrimidin-2-yl)piperazine-1-carboxylate (7.5 g, yield
60%) as a white solid, MS (ES+) C.sub.20H.sub.26N.sub.4O.sub.2
requires: 354. found: 355 [M+H].sup.+.
Step 7: Synthesis of 5-benzyl-2-(piperazin-1-yl)pyrimidine Hydrogen
Chloride Salt
##STR00315##
To a solution of tert-butyl
4-(5-benzylpyrimidin-2-yl)piperazine-1-carboxylate (8.0 g, 23.0
mmol) in 1,4-dioxane (30 mL) was dropwise added a solution of 4 M
HCl-dioxane (20 mL, 80 mmol). The reaction mixture was stirred at
RT overnight, and then concentrated to give
5-benzyl-2-(piperazin-1-yl)pyrimidine HCl salt (10.2 g, crude) as a
yellow solid, MS (ES+) C.sub.15H.sub.18N.sub.4 requires: 254.
found: 255 [M+H-100].sup.+.
Step 8: Synthesis of
2-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-4-chloro-1,3,5-triazine
##STR00316##
A solution of 2,4-dichloro-1,3,5-triazine (315 mg, 2.1 mmol),
5-benzyl-2-(piperazin-1-yl)pyrimidine (510 mg, 2.0 mmol) and
Hunig's base (540 mg, 4.2 mmol) in 1,4-dioxane (6 mL) was stirred
at 60.degree. C. for 1 h. LCMS showed the reaction was completed.
The reaction mixture was cooled to RT, diluted by water (60 mL),
and extracted with ethyl acetate (40 mL.times.3). The organic
layers were combined, washed with water and brine, dried over
sodium sulfate, filtered and concentrated to give
1-(2,2-dimethyl-1,3-dioxolan-4-yl)-1H-pyrazol-4-amine (280 mg,
crude) as a yellow solid, which was directly used in the next step
without further purification. MS (ES+) C.sub.18H.sub.18ClN.sub.7
requires: 367. found: 368 [M+H].sup.+.
Step 9: Synthesis of
4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-N-(1-((2,2-dimethyl-1,3-diox-
olan-4-yl)methyl)-1H-pyrazol-4-yl)-1,3,5-triazin-2-amine
##STR00317##
A solution of
2-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-4-chloro-1,3,5-triazine
(110 mg, 0.3 mmol),
1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-pyrazol-4-amine (90
mg, 0.45 mmol) and Hunig's base (80 mg, 0.6 mmol) in 1,4-dioxane (4
mL) was stirred at 60.degree. C. for 5 h. LCMS showed the reaction
was completed. The reaction mixture was cooled to RT and diluted
with ethyl acetate (40 mL). The organic phase was washed with water
(40 mL) and brine (20 mL), dried over sodium sulfate, filtered and
concentrated to give
4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-N-(1-((2,2-dimethyl-1,3-diox-
olan-4-yl)methyl)-1H-pyrazol-4-yl)-1,3,5-triazin-2-amine (200 mg,
crude) as a yellow solid, which was directly used in the next step
without further purification. MS (ES+)
C.sub.26H.sub.30N.sub.10O.sub.2 requires: 528. found: 529
[M+H].sup.+.
Step 10: Synthesis of
3-(4-(4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylamin-
o)-1H-pyrazol-1-yl)propane-1,2-diol
##STR00318##
A solution of
4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-N-(1-((2,2-dimethyl-1,3-diox-
olan-4-yl)methyl)-1H-pyrazol-4-yl)-1,3,5-triazin-2-amine (200 mg,
0.3 mmol), and p-TSA (40 mg, 0.2 mmol) in dichloroethane (2.5 mL)
was stirred at 80.degree. C. under N.sub.2 overnight. LCMS showed
the reaction was completed. The reaction mixture was cooled to RT
and diluted with ethyl acetate (40 mL). The organic phase was
washed with water (30 mL) and brine (30 mL), dried over sodium
sulfate, filtered and concentrated. The residue was purified by
Prep-HPLC to give
3-(4-(4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylamin-
o)-1H-pyrazol-1-yl)propane-1,2-diol (13.8 mg, yield 7%) as a white
solid. MS (ES+) C.sub.24H.sub.28N.sub.10O.sub.2 requires: 488.
found: 489 [M+H].sup.+.
##STR00319## ##STR00320##
Scheme 4 schematically depicts synthetic protocol 4. The piperazine
carbonyl derivative, e.g., carbamoyl, (A, X and Y are each --CH--)
can be coupled to the Grignard bromide (2, Ring A is aryl), to
provide the protected di-substituted carbonyl (C, X.sup.1 is
CH.sub.2, S, NH, or O). When X.sup.1 is O, i.e., a carbonyl, the
carbonyl can be further reacted with an organometallic reagent such
as trialkylaluminum, e.g., trimethylaluminum, which can also
deprotect the piperazine nitrogen to provide the dialkyl compound
(C', alkyl is C.sub.1-6 alkyl). Removal of the protecting group (P)
from the piperazine ring of (C) can be carried out using strong
acids such as 4M hydrochloric acid (HCl) in dioxane or
trifluoroacetic acid (TFA) in a polar solvent such as methanol or
dichloromethane (DCM) to afford amine (D). Triazine (E) can be
substituted with amine (C') or (D) under nucleophilic aromatic
substitution reaction conditions using an amine base such as
diisopropylethylamine (DIPEA) or triethylamine (TEA) in a polar
solvent such as dioxane to provide the piperazine-substituted
triazine (F) or (F'). Reduction of --C(.dbd.X.sup.1)--, wherein
X.sup.1 is CH.sub.2, S, NH, or O, e.g., carbonyl, of (F) can be
performed using a reducing agent such as sodium borohydride to
provide --C--(XH)--, e.g., the alcohol (G). As shown below,
Compound 189 was prepared using synthetic protocol 4.
Synthesis of (S) (2 (4 (4 (1
(morpholin-2-ylmethyl)-1H-pyrazol-4-ylamino)-1,3,5-triazin-2-yl)piperazin-
-1-yl)pyrimidin-5-yl)(phenyl)methanol (Compound 189)
##STR00321## ##STR00322##
Step 1: Synthesis of ethyl
2-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrimidine-5-carboxylate
##STR00323##
To a solution of tert-butyl piperazine-1-carboxylate (7.9 g, 42.5
mmol) and diisopropylethylamine (13.69 g, 106.1 mmol) in
dichloromethane (80 mL) was added ethyl
2-chloropyrimidine-5-carboxylate (7.9 g, 42.5 mmoL), and the
reaction mixture was stirred at room temperature for 3 hours. LCMS
showed the reaction was completed. The reaction mixture was
directly concentrated to afford ethyl
2-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrimidine-5-carboxylate
(17 g, crude), which was directly used in the next step without
further purification. MS (ES+) C.sub.16H.sub.24N.sub.4O.sub.4
requires: 336. found: 237, 281 [M-56+H].sup.+.
Step 2: Synthesis of
2-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrimidine-5-carboxylic
acid
##STR00324##
To a solution of ethyl
2-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrimidine-5-carboxylate
(17 g, crude) in THF/methanol/H.sub.2O (300 mL) was added sodium
hydroxide (4.3 g, 107.5 mmol), and the reaction mixture was stirred
at 70 0.degree. C. for 2 hours. LCMS showed the reaction was
completed. The reaction mixture was brought to pH.apprxeq.5-6 with
1 M HCl, and then filtered. The solid was collected and dried to
give
2-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrimidine-5-carboxylic
acid as a white solid (16 g, 96%), which was directly used in the
next step without further purification. MS (ES+)
C.sub.14H.sub.20N.sub.4O.sub.4 requires: 308. found: 253
[M-56+H].sup.+.
Step 3: Synthesis of tert-butyl
4-(5-(methoxy(methyl)carbamoyl)pyrimidin-2-yl)piperazine-1
##STR00325##
To a suspension of
2-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrimidine-5-carboxylic
acid (13.8 g, 44.8 mmol),
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (12.8 g, 67.2 mmol)
and hydroxybenzotriazole (7.2 g, 53.7 mmol) in dichloromethane (200
mL) was added triethylamine (25 mL, 179.2 mmol). The mixture was
stirred at room temperature for 1 hour, and then
N,O-dimethylhydroxylamine (5 g, 53.7 mmol) was added. The reaction
mixture was stirred for another 3 hours. LCMS showed the reaction
was completed. The reaction mixture was directly washed with water
(100 mL), and the organic layer was dried over sodium sulfate,
filtered and concentrated. The residue was purified by silica gel
chromatography (petroleum ether:ethyl acetate=1:1) to give
tert-butyl
4-(5-(methoxy(methyl)carbamoyl)pyrimidin-2-yl)piperazine-1-carboxylate
(11.2 g, 67%) as a white solid. MS (ES+)
C.sub.16H.sub.25N.sub.5O.sub.4 requires: 351. found: 296
[M-56+H].sup.+.
Step 4: Synthesis of tert-butyl
4-(5-benzoylpyrimidin-2-yl)piperazine-1-carboxylate
##STR00326##
To a solution of tert-butyl
4-(5-(methoxy(methyl)carbamoyl)pyrimidin-2-yl)piperazine-1-carboxylate
(2.6 g, 7.4 mmol) in dry THF (30 mL) was added benzylmagnesium
bromide (1 M in THF, 29.6 mL) at 0.degree. C. under N.sub.2, and
the mixture was stirred at room temperature for 3 hours. LCMS
showed the reaction was completed. The reaction mixture was
quenched with 1 M HCl and extracted with ethyl acetate. The organic
layer was washed with water and brine, dried over sodium sulfate,
filtered and concentrated. The residue was purified by silica gel
chromatography (petroleum ether:ethyl acetate=5:1) to get
tert-butyl 4-(5-benzoylpyrimidin-2-yl)piperazine-1-carboxylate (2
g, 73%) as a yellow solid. MS (ES+) C.sub.20H.sub.24N.sub.4O.sub.3
requires: 368. found: 313 [M-56+H].sup.+.
Step 5: Synthesis of
phenyl(2-(piperazin-1-yl)pyrimidin-5-yl)methanone HCl salt
##STR00327##
To a solution of tert-butyl
4-(5-benzoylpyrimidin-2-yl)piperazine-1-carboxylate (1 g, 2.7 mmol)
in dioxane (20 mL) was added 4 M HCl-dioxane (20 mL), and the
reaction mixture was stirred at room temperature overnight. LCMS
showed the reaction was completed. The mixture was directly
concentrated to give
phenyl(2-(piperazin-1-yl)pyrimidin-5-yl)methanone HCl salt as a
yellowish solid (0.95 g, 90%). MS (ES+) C.sub.15H.sub.16N.sub.4O
requires: 268. found: 269 [M+H].sup.+.
Step 6: Synthesis of (S)-tert-butyl
2-((4-(4-(4-(5-benzoylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylam-
ino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
##STR00328##
The synthesis of (S)-tert-butyl
2-((4-(4-chloro-1,3,5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholin-
e-4-carboxylate (7) was carried out following the synthetic
procedure of Example 1.)
To a mixture of phenyl(2-(piperazin-1-yl)pyrimidin-5-yl)methanone
HCl salt (141.6 mg, 0.38 mmol) and (S)-tert-butyl
2-((4-(4-chloro-1,3,5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholin-
e-4-carboxylate (150 mg, 0.38 mmol) in dioxane (10 mL) was added
diisopropylethylamine (1 mL), and the reaction was stirred at room
temperature overnight. LCMS showed the reaction was completed. The
solvents were removed under reduced pressure, and the residue was
purified by silica gel chromatography (petroleum ether:ethyl
acetate=1:1) to give (S)-tert-butyl
2-((4-(4-(4-(5-benzoylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylam-
ino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate (230 mg, 95%)
as a yellowish solid. (MS (ES+) C.sub.31H.sub.37N.sub.11O.sub.4
requires: 627. found: 573 [M-56+H].sup.+.
Step 7: Synthesis of (S)-tert-butyl
2-((4-(4-(4-(5-(hydroxy(phenyl)methyl)pyrimidin-2-yl)piperazin-1-yl)-1,3,-
5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
##STR00329##
To a solution of (S)-tert-butyl
2-((4-(4-(4-(5-benzoylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylam-
ino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate (80 mg, 0.127
mmol) in THF/methanol was added sodium borohydride (20 mg, 0.51
mmol). The mixture was stirred at room temperature for 1 hour, and
LCMS showed the reaction was completed. The solvents were removed
to give (S)-tert-butyl
2-((4-(4-(4-(5-(hydroxy(phenyl)methyl)pyrimidin-2-yl)piperazin-1-yl)-1,3,-
5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
(80 mg, crude) as a yellowish solid. (MS (ES+)
C.sub.31H.sub.39N.sub.11O.sub.4 requires: 629. found: 630
[M+H].sup.+.
Step 8: Synthesis of
(S)-(2-(4-(4-(1-(morpholin-2-ylmethyl)-1H-pyrazol-4-ylamino)-1,3,5-triazi-
n-2-yl)piperazin-1-yl)pyrimidin-5-yl)(phenyl)methanol
##STR00330##
To a solution of (S)-tert-butyl
2-((4-(4-(4-(5-benzoylpyrimidin-2-yl)piperazin-1-yl)-1,3,5-triazin-2-ylam-
ino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate (80 mg, 0.127
mmol, crude) in dioxane (4 mL) was added 4 M HCl-dioxane (4 mL),
and the reaction was stirred at room temperature overnight. LCMS
showed the reaction was completed. The solvent was removed, and the
residue was purified by Prep-HPLC to provide the title compound
(S)-(2-(4-(4-(1-(morpholin-2-ylmethyl)-1H-pyrazol-4-ylamino)-1,3,5-triazi-
n-2-yl)piperazin-1-yl)pyrimidin-5-yl)(phenyl)methanol (Compound
189) (24.0 mg, 18%) as a yellowish solid.
##STR00331## ##STR00332##
Scheme 5 schematically depicts synthetic protocol 5. The
N-protected heteroaryl-substituted piperazine (A, X and Y are each
--CH--) can be coupled to an alcohol, thiol, or amine, (B, --Z--H
is --OH, --SH, or --NH.sub.2; Ring A is aryl) via a copper-mediated
coupling reaction, e.g., such as the Ullman reaction, to provide
the protected heteroaryl-ether (C). Removal of the protecting group
(P) from the piperazine ring can be carried out using strong acids
such as 4M hydrochloric acid (HCl) in dioxane or trifluoroacetic
acid (TFA) in a polar solvent such as methanol or dichloromethane
(DCM) to afford amine (D). Triazine (E) can be substituted with
amine (D) under nucleophilic aromatic substitution reaction
conditions using an amine base such as diisopropylethylamine
(DIPEA) or triethylamine (TEA) in a polar solvent such as dioxane
to provide the piperazine-substituted triazine (F). As shown below,
Compound 203 was prepared using synthetic protocol 5.
Synthesis of
(S)-4-(4-(5-(2-fluorophenoxy)pyrimidin-2-yl)piperazin-1-yl)-N-(1-(morphol-
in-2-ylmethyl)-1H-pyrazol-4-yl)-1,3,5-triazin-2-amine (Compound
203)
##STR00333## ##STR00334##
Step 1: Synthesis of tert-butyl
4-(5-(2-fluorophenoxy)pyrimidin-2-yl)piperazine-1-carboxylate
##STR00335##
A mixture of tert-butyl
4-(5-bromopyrimidin-2-yl)piperazine-1-carboxylate (684 mg, 2.0
mmol), 2-fluorophenol (1.1 g, 10.0 mmol), copper (650 mg, 10.0
mmol) and cesium carbonate (6.5 g, 20.0 mmol) in pyridine (15 mL)
was heated at 120 0.degree. C. in a sealed tube overnight. The
reaction mixture was diluted with ethyl acetate (200 mL) and then
filtered. The filtrate was concentrated, and the residue was
purified by silica gel chromatography (petroleum ether:ethyl
acetate 20:1) to give tert-butyl
4-(5-(2-fluorophenoxy)pyrimidin-2-yl)piperazine-1-carboxylate (50
mg, 6%) as a yellow solid. MS (ES+) C.sub.19H.sub.23FN.sub.4O.sub.3
requires: 374. found: 397 [M+Na].sup.+.
Step 2: Synthesis of
5-(2-fluorophenoxy)-2-(piperazin-1-yl)pyrimidine HCl Salt
##STR00336##
To a solution of tert-butyl
4-(5-(2-fluorophenoxy)pyrimidin-2-yl)piperazine-1-carboxylate (50
mg, 0.13 mmol) in dioxane (3 mL) was added 4 M HCl/dioxane (3 mL).
The reaction mixture was stirred at room temperature overnight and
then concentrated to dryness. The residue (50 mg, yellow solid) was
directly used in the next reaction.
Step 3: Synthesis of (S)-tert-butyl
2-((4-(4-(4-(5-(2-fluorophenoxy)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-tria-
zin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
##STR00337##
The synthesis of (S)-tert-butyl
2-((4-(4-chloro-1,3,5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholin-
e-4-carboxylate (5) was carried out following the synthetic
procedure of Example 1.
A mixture of 5-(2-fluorophenoxy)-2-(piperazin-1-yl)pyrimidine HCl
salt (27 mg, 0.1 mmol), (S)-tert-butyl
2-((4-(4-chloro-1,3,5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)-morpholi-
ne-4-carboxylate (40 mg, 0.1 mmol) and diisopropylethylamine (25
mg, 2.0 mmol) in dioxane (2 mL) was stirred at room temperature for
2 hours. The reaction mixture was directly evaporated and used in
the next step.
Step 4: Synthesis of
(S)-4-(4-(5-(2-fluorophenoxy)pyrimidin-2-yl)piperazin-1-yl)-N-(1-(morphol-
in-2-ylmethyl)-1H-pyrazol-4-yl)-1,3,5-triazin-2-amine
##STR00338##
The above crude sample was dissolved in dichloromethane (10 mL),
followed by the addition of trifluoroacetic acid (4 mL). The
mixture was stirred at room temperature overnight. The solvent was
removed, and the residue was purified by Prep-HPLC to
(S)-4-(4-(5-(2-fluorophenoxy)-pyrimidin-2-yl)piperazin-1-yl)-N-(1-(morpho-
lin-2-ylmethyl)-1H-pyrazol-4-yl)-1,3,5-triazin-2-amine (4.3 mg) as
a white solid.
##STR00339## ##STR00340##
Scheme 6 schematically depicts synthetic protocol 6. The
N-protected heteroaryl-substituted piperazine (A, X and Y are each
--CH--) can be coupled to alkyne (B, Ring A is cycloalkyl) via a
palladium-mediated coupling reaction, e.g., Sonogashira coupling,
to provide the protected heteroaryl-alkyne (C). Removal of the
protecting group (P) from the piperazine ring can be carried out
using strong acids such as 4M hydrochloric acid (HCl) in dioxane or
trifluoroacetic acid (TFA) in a polar solvent such as methanol or
dichloromethane (DCM) to afford amine (D). Triazine (E) can be
substituted with amine (D) under nucleophilic aromatic substitution
reaction conditions using an amine base such as
diisopropylethylamine (DIPEA) or triethylamine (TEA) in a polar
solvent such as dioxane to provide the piperazine-substituted
triazine (F). As shown below, Compound 63 was prepared using
synthetic protocol 6.
Synthesis of
(S)-4-(4-(5-(cyclopropylethynyl)pyrimidin-2-yl)piperazin-1-yl)-N-(1-(morp-
holin-2-ylmethyl)-1H-pyrazol-4-yl)-1,3,5-triazin-2-amine (Compound
63)
##STR00341## ##STR00342##
Step 1: Synthesis of tert-butyl
4-(5-(cyclopropylethynyl)pyrimidin-2-yl)piperazine-1-carboxylate
##STR00343##
In a sealed tube, the mixture of tert-butyl
4-(5-bromopyrimidin-2-yl)piperazine-1-carboxylate (1.0 g, 2.9
mmol), ethynylcyclopropane (482 mg, 7.3 mmol),
tetrakis(triphenyl-phosphine)palladium (335 mg, 0.29 mmol) and
copper(I) iodide (28 mg, 0.15 mmol) in diethylamine (10 mL) and
dimethyl sulfoxide (10 mL) was stirred at 100.degree. C. for 3
hours under nitrogen atmosphere. The mixture was then diluted with
ethyl acetate (100 mL) and washed with water and brine. The organic
phase was dried over sodium sulfate, filtered and concentrated. The
residue was purified by silica gel chromatography eluting with
petroleum ether:ethyl acetate=15:1 to afford tert-butyl
4-(5-(cyclopropylethynyl)pyrimidin-2-yl)piperazine-1-carboxylate
(930 mg, 98%) as a yellow solid. MS (ES+)
C.sub.18H.sub.24N.sub.4O.sub.2 requires: 328. found: 329
[M+H].sup.+.
Step 2: Synthesis of
5-(cyclopropylethynyl)-2-(piperazin-1-yl)pyrimidine HCl salt
##STR00344##
To a solution of tert-butyl
4-(5-(cyclopropylethynyl)pyrimidin-2-yl)piperazine-1-carboxylate
(150 mg, 0.46 mmol) in dioxane (3 mL) was added 4 M HCl-dioxane (3
mL), and the reaction mixture was stirred room temperature for 1
hour. LCMS showed the reaction was completed. The reaction mixture
was concentrated to afford
5-(cyclopropylethynyl)-2-(piperazin-1-yl)pyrimidine HCl salt as a
white solid (100 mg, crude), which was directly used in the next
step without further purification. MS (ES+) C.sub.13H.sub.16N.sub.4
requires: 228. found: 229 [M+H].sup.+.
Step 3: Synthesis of (S)-tert-butyl
2-((4-(4-(4-(5-(cyclopropylethynyl)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-t-
riazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
##STR00345##
The synthesis of (S)-tert-butyl
2-((4-(4-chloro-1,3,5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholin-
e-4-carboxylate 5 was performed as shown elsewhere herein.
To a solution of
5-(cyclopropylethynyl)-2-(piperazin-1-yl)pyrimidine HCl salt (172
mg, 0.44 mmol) and (S)-tert-butyl
2-((4-(4-chloro-1,3,5-triazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholin-
e-4-carboxylate was added diisopropylethylamine (169 mg, 1.311
mmol), and the reaction mixture was stirred at room temperature.
LCMS showed the reaction was completed. The mixture was
concentrated, and the residue was purified by silica gel
chromatography, eluting with petroleum ether: ethyl acetate=1:2, to
afford (S)-tert-butyl
2-((4-(4-(4-(5-(cyclopropylethynyl)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-t-
riazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
(240 mg, 94%) as a yellowish solid. MS (ES+):
C.sub.29H.sub.37N.sub.11O.sub.3 requires: 587. found: 588
[M+H].sup.+.
Step 4: Synthesis of
(S)-4-(4-(5-(cyclopropylethynyl)pyrimidin-2-yl)piperazin-1-yl)-N-(1-(morp-
holin-2-ylmethyl)-1H-pyrazol-4-yl)-1,3,5-triazin-2-amine
##STR00346##
To a solution of (S)-tert-butyl
2-((4-(4-(4-(5-(cyclopropylethynyl)pyrimidin-2-yl)piperazin-1-yl)-1,3,5-t-
riazin-2-ylamino)-1H-pyrazol-1-yl)methyl)morpholine-4-carboxylate
(30 mg, 0.05 mmol) in dichloromethane (2 mL) was dropwise added
trifluoroacetic acid (1 mL). The reaction mixture was stirred at
room temperature for 0.5 hour and concentrated. The residue was
purified by Prep-HPLC to afford
(S)-4-(4-(5-(cyclopropylethynyl)pyrimidin-2-yl)piperazin-1-yl)-N-(1-(morp-
holin-2-ylmethyl)-1H-pyrazol-4-yl)-1,3,5-triazin-2-amine (14 mg,
57%) as a white solid.
Preparation of Common Intermediates
Synthesis of
5-(2-phenylpropan-2-yl)-2-(piperazin-1-yl)pyrimidine
##STR00347##
In a sealed tube, the mixture of tert-butyl
4-(5-benzoylpyrimidin-2-yl) piperazine-1-carboxylate (500 mg, 1.36
mmol) and trimethylaluminum (2 M in toluene, 2.7 mL) in dry toluene
(10 mL) was stirred at 100 0.degree. C. overnight. LCMS showed the
reaction was completed. The reaction mixture was cooled to RT,
quenched with ice-water and extracted with ethyl acetate. The
organic layer was washed with water and brine, dried over sodium
sulfate, filtered and concentrated. The residue was purified by
Prep-HPLC to get
5-(2-phenylpropan-2-yl)-2-(piperazin-1-yl)pyrimidine (40 mg, 7%) as
a yellowish solid. MS (ES+) C.sub.17H.sub.22N.sub.4 requires: 282.
found: 283 [M+H].sup.+.
Synthesis of 2-methyl-1-(4-nitro-1H-pyrazol-1-yl)propan-2-ol
##STR00348##
A mixture of 4-nitro-1H-pyrazole (1.6 g, 13.9 mmol),
1-chloro-2-methylpropan-2-ol (1.5 g, 13.9 mmol) and
Cs.sub.2CO.sub.3 (9.1 g, 27.8 mmol) in DMF (20 mL) was stirred at
80.degree. C. for 5 h. The reaction mixture was cooled to RT and
diluted with EA (200 ml). The organic phase was washed by water (50
mL) and brine (50 mL), dried over anhydrous sodium sulfate and
concentrated under reduced pressure to give
2-methyl-1-(4-nitro-1H-pyrazol-1-yl)propan-2-ol as a yellow oil
(2.0 g, 78%), which was used for the next step without further
purification. MS (ES+) C.sub.7H.sub.11N.sub.3O.sub.3 requires: 185.
found: 186 [M+H].sup.+.
Synthesis of 1-(4-amino-1H-pyrazol-1-yl)-2-methylpropan-2-ol
##STR00349##
A stirred suspension mixture of
2-methyl-1-(4-nitro-1H-pyrazol-1-yl)propan-2-ol (2.0 g, 10.8 mmol)
and 10% Pd/C (0.2 g, 0.1 w/w) in MeOH (20 mL) was exposed to 1 atm
H.sub.2 at RT overnight. The reaction mixture was filtered through
a pad of celite. The filtrate was concentrated under reduced
pressure to give 1-(4-amino-1H-pyrazol-1-yl)-2-methylpropan-2-ol as
a yellow oil (1.9 g, 99%), which was used for the next step without
further purification. MS (ES+) C.sub.7H.sub.13N.sub.3O requires:
155. found: 156 [M+H].sup.+.
Synthesis of 1-((4-bromophenyl)diazenyl)pyrrolidine
##STR00350##
To a solution of 4-bromoaniline (1.7 g, 9.88 mmol) in conc. HCl (2
mL) was added a solution of NaNO.sub.2 (682 mg, 9.88 mmol) in water
(3 mL) at 0.degree. C. The solution was stirred at 0.degree. C. for
10 minutes, followed by the addition of pyrrolidine (844 mg, 11.86
mmol) in KOH solution (1 mL, 1 N). The resultant mixture was
stirred at 0.degree. C. for 0.5 h. The formed precipitate was
collected by filtration, washed with 1 mL of ethanol and dried to
give the title compound as a yellow solid (1.5 g, yield 60%). MS
(ES+) C.sub.10H.sub.12BrN.sub.3 requires: 253, 255. found 254, 256
[M+H].sup.+, purity: 93%.
Synthesis of 3-(4-(pyrrolidin-1-yldiazenyl)phenyl)oxetan-3-ol
##STR00351##
To a solution of 1-((4-bromophenyl)diazenyl)pyrrolidine (500 mg,
1.97 mmol) in anhydrous THF (20 mL) was added n-BuLi (1.8 mL, 4.33
mmol) dropwise at -78.degree. C. under N.sub.2. The solution was
stirred at -78.degree. C. for 1 h, followed by the addition of
oxetan-3-one (326 mg, 4.53 mmol). The resultant mixture was stirred
at RT for 18 h. The reaction was quenched by saturated aqueous
NH.sub.4Cl aq. (20 mL) and extracted with ethyl acetate (20 mL*3).
The combined organic layers were concentrated under reduced
pressure to give a residue, which was purified by silica gel
chromatography (DCM:MeOH=20:1) to afford the title compound (500
mg, yield 97%) as a light yellow solid. MS (ES+)
C.sub.13H.sub.17N.sub.3O.sub.2 requires: 247. found 248
[M+H].sup.+, purity: 90%.
Synthesis of 3-(4-aminophenyl)oxetan-3-ol
##STR00352##
To a solution of 3-(4-(pyrrolidin-1-yldiazenyl)phenyl)oxetan-3-ol
(500 mg, 2.02 mmol) in methanol (15 mL) was added Pd/C (10%, 250
mg) at RT. The reaction mixture was stirred under 1 atm H.sub.2
atmosphere (balloon) at 40.degree. C. for 18 h. The mixture was
filtered through a pad of Celite. The filtrate was concentrated to
give a residue, which was purified by Prep-HPLC to afford the title
compound (200 mg, yield 60%) as a white solid. MS (ES+)
C.sub.9H.sub.11NO.sub.2 requires: 165. found 166 [M+H].sup.+,
purity: 96%.
Synthesis of tert-butyl 1-(2-methylprop-1-enyl)-4-nitrobenzene
##STR00353##
A mixture of 1-iodo-4-nitrobenzene (2.49 g, 10.0 mmol),
4,4,5,5-tetramethyl-2-(2-methylprop-1-enyl)-1,3,2-dioxaborolane
(1.82 g, 10.0 mmol), sodium bicarbonate (2.52 g, 30.0 mmol) and
[1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II) complex
with dichloromethane (816 mg, 1.0 mmol) in dioxane/water (5:1, 60
mL) was stirred at 90.degree. C. for 20 h under N.sub.2. LCMS and
TLC monitored the reaction was completed. The reaction mixture was
cooled to RT and concentrated. The residue was dissolved in 100 mL
of EtOAc, washed with water (150 mL.times.3) and brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under vacuo. The crude
product was purified by silica gel column, eluting with PE:EA
(6:1), to obtain the desired product (1.9 g, 100% yield) as a
yellow oil, .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.17 (d,
2H, J=8.8 Hz), 7.35 (d, 2H, J=8.8 Hz), 6.36 (s, 1H), 1.96 (d, 3H,
J=0.4 Hz), 1.90 (d, 3H, J=0.4 Hz).
Synthesis of 2-methyl-1-(4-nitrophenyl)propane-1,2-diol
##STR00354##
To a solution of N-methyl morpholine-N-oxide (1.5 g, 12.8 mmol) in
4.5 mL of water was added a solution of tert-butyl
1-(2-methylprop-1-enyl)-4-nitrobenzene (1.5 g, 8.5 mmol) in
acetone/water (5:1, 36 mL), followed by the addition of a solution
of osmium tetraoxide in water (2 mL, 4%). This mixture was allowed
to RT and stirred for 16 h. The reaction was quenched by sat.
Na.sub.2SO.sub.3. aq (100 mL) and extracted with EtOAc (50
mL.times.3). The combined organic layers were washed with water and
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated under
vacuo. The crude product (1.7 g, yield 100%) was obtained as a
yellow foam, which was directly used into the next step without
further purification. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. ppm
8.20 (d, 2H, J=9.2 Hz), 7.57 (d, 2H, J=9.2 Hz), 4.63 (d, 1H, J=2.4
Hz), 2.69 (d, 1H, J=3.2 Hz), 2.03 (d, 1H, J=3.2 Hz), 1.19 (s, 3H),
1.08 (s, 3H).
Synthesis of
2,2,4,4-tetramethyl-5-(4-nitrophenyl)-1,3-dioxolane
##STR00355##
A solution of 2-methyl-1-(4-nitrophenyl)propane-1,2-diol (1.5 g,
7.1 mmol), 2,2-dimethoxypropane (1.45 g, 14.2 mmol) and TsOH (cat.,
260 mg, 1.5 mmol) in acetone (40 mL) was stirred at 30.degree. C.
for 16 h. The reaction mixture was concentrated. The residue was
purified by silica gel chromatography, eluting with PE:EA (8:1), to
obtain the desired product (1.5 g, 85% yield) as a white solid.
.sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. ppm 8.23 (d, 2H, J=9.0
Hz), 7.57 (d, 2H, J=9.0 Hz), 4.93 (s, 1H), 1.59 (s, 3H), 1.48 (s,
3H), 1.46 (s, 3H), 0.78 (s, 3H).
Synthesis of (R) and
(S)-2,2,4,4-tetramethyl-5-(4-nitrophenyl)-1,3-dioxolane
##STR00356##
The racemic material
2,2,4,4-tetramethyl-5-(4-nitrophenyl)-1,3-dioxolane (1.3 g) was
separated by chiral HPLC to obtain peak 1 (630 mg, 100% ee) and
peak 2 (610 mg, 99.6% ee).
Synthesis of
(R)-4-(2,2,5,5-tetramethyl-1,3-dioxolan-4-yl)aniline
##STR00357##
A mixture of
(R)-2,2,4,4-tetramethyl-5-(4-nitrophenyl)-1,3-dioxolane (200 mg,
0.797 mmol) and Pd/C (10%, 60 mg) in EtOAc/IPA (10 mL/10 mL) was
stirred under 1 atm H.sub.2 atmosphere (balloon) at RT for 16 h.
The mixture was filtered through a pad of Celite. The filtrate was
concentrated under reduced pressure to afford the desired product
(180 mg, 100%), which was directly used into the next step without
further purification.
Synthesis of 1-(4-nitrophenyl)ethane-1,2-diol
##STR00358##
To a stirred solution of ethyl 2-(4-nitrophenyl)-2-oxoacetate (10.0
g, 47.8 mmol) in ethanol (100 mL) was slowly added sodium
borohydride (4.54 g, 119.5 mmol) at 0.degree. C. After the addition
was completed, the reaction mixture was allowed to warm to RT and
stirred for 12 h. The reaction was quenched by acetone (10 mL) and
concentrated. The residue was purified by silica gel column
chromatography (dichloromethane/methanol=5/1) to afford the title
compound (8.0 g, 87%) as a brown solid. MS (ES+)
C.sub.8H.sub.9NO.sub.4 requires: 183. found: 184 [M+H].sup.+.
Synthesis of 2,2-dimethyl-4-(4-nitrophenyl)-1,3-dioxolane
##STR00359##
A mixture of 1-(4-nitrophenyl)ethane-1,2-diol (7.0 g, 38.3 mmol),
2,2-dimethoxypropane (16 g, 153.2 mmol) and 4-methylbenzenesulfonic
acid (2.6 g, 15.32 mmol) in acetone (100 mL) was stirred at
25.degree. C. for 12 h. The reaction mixture was concentrated and
then dissolved in ethyl acetate (300 mL). The organic layer was
separated, washed by water (100 mL) and brine (100 mL), dried over
sodium sulfate, filtered and concentrated. The residue was purified
by silica gel column chromatography (petroleum ether/ethyl
acetate=5/1) to afford the title compound (4.0 g, 40%) as a yellow
oil.
The racemic material was separated by chiral-HPLC to afford the
desired isomer (shown on the bottom, above) (1.8 g, 45%).
Synthesis of (S)-4-(2,2-dimethyl-1,3-dioxolan-4-yl)benzenamine
##STR00360##
A mixture of (S)-2,2-dimethyl-4-(4-nitrophenyl)-1,3-dioxolane (3.6
g, 16.1 mmol) and 5% palladium on carbon (50% wet, 700 mg) in
methanol (30 mL) was stirred under 1 atm hydrogen atmosphere
(H.sub.2 balloon) at RT for 16 h. After the reaction mixture was
filtered through a pad of Celite, the filtrate was concentrated to
afford the title compound (3.1 g, 100%) as a yellow solid. MS (ES+)
C.sub.11H.sub.15NO.sub.2 requires: 193. found: 194 [M+H].sup.+.
Synthesis of tert-butyl
4-(5-acetylpyrimidin-2-yl)piperazine-1-carboxylate
##STR00361##
To a mixture of tert-butyl
4-(5-bromopyrimidin-2-yl)piperazine-1-carboxylate (5.0 g, 14.6
mmol), palladium diacetate (240 mg, 1.46 mmol) and
triphenylphosphine (376 mg, 2.92 mmol) in dioxane (100 mL) was
added tributyl(1-ethoxyvinyl)stannane (5.3 mL, 16.1 mL) under
N.sub.2, and the reaction mixture was stirred at 80.degree. C.
overnight. The reaction was cooled to RT and diluted with THF (100
mL), followed by the addition of 2 N HCl (100 mL). The mixture was
stirred at RT for 30 mins, and LCMS showed the reaction was
completed. The reaction mixture was diluted with ethyl acetate (200
mL). The organic phase was separated, washed with water
(3.times.100 mL), dried over sodium sulfate, filtered and
concentrated. The residue was purified by silica gel column
chromatography to afford the title compound (3.0 g, 67%). MS (ES+)
C.sub.15H.sub.22N.sub.4O.sub.3 requires: 306. found: 251
[M-56+H].sup.+.
Synthesis of 1-(2-(piperazin-1-yl)pyrimidin-5-yl)ethanone
##STR00362##
To a solution of -butyl
4-(5-acetylpyrimidin-2-yl)piperazine-1-carboxylate (4.5 g, 14.7
mmol) in dichloromethane (80 mL) was added trifluoroethyl acetate
(20 mL), and the mixture was stirred at RT for 3 h. LCMS showed the
reaction was completed. The reaction mixture was neutralized with
sodium carbonate solution and extracted with dichloromethane. The
organic layer was dried over sodium sulfate, filtered and
concentrated to afford the title compound as a light yellow solid
(2.6 g, 86%), which was directly used in the next step without
further purification. MS (ES+) C.sub.10H.sub.14N.sub.4O requires:
206. found: 207 [M+H].sup.+.
Synthesis of
1-(4-fluorophenyl)-1-(2-(piperazin-1-yl)pyrimidin-5-yl)ethanol
##STR00363##
To a solution of 1-(2-(piperazin-1-yl)pyrimidin-5-yl)ethanone (1.8
g, 8.73 mmol) in dry THF (100 mL) was added
(4-fluorophenyl)magnesium bromide (1 M in THF, 87.3 mL) at
0.degree. C. under N.sub.2. The mixture was stirred at RT for 3 h,
then quenched with ammonium chloride solution and extracted with
dichloromethane (300 mL). The organic layer was dried over sodium
sulfate, filtered and concentrated. The residue was purified by
Combi-flash (dicholomethane:methanol=10:1) to give the title
compound (1.02 g, 38%) as a yellow solid.
Chiral separation of
1-(4-fluorophenyl)-1-(2-(piperazin-1-yl)pyrimidin-5-yl)ethanol
##STR00364##
The racemate compound (1.02 g) was separated by chiral HPLC to
afford enantiomer 1 (320 mg) and enantiomer 2 (220 mg). MS (ES+)
C.sub.16H.sub.19FN.sub.4O requires: 302. found: 303 [M+H].sup.+.
The absolute configuration was assigned randomly.
Chiral separation conditions: Chiral column: OZ--H (4.6*250 mm,
Sum); Mobile phase: co-solvent EtOH (0.1% DEA)
Synthesis of (4-nitrophenyl)methanesulfonamide
##STR00365##
To a solution of (4-nitrophenyl)methanesulfonyl chloride (4 g, 17
mmol) in acetonitrile (100 mL) was added a solution of ammonium
carbonate (3 g, 31.3 mmol) in ammonia (50 mL). The reaction mixture
was stirred for 1 h, and TLC indicated the starting material
consumed completely. The organic solvent was removed under reduced
pressure. Addition of water (10 mL) led to precipitate formation.
The solid was collected, washed with water (2.times.5 mL), and
dried to give (4-nitrophenyl)methanesulfonamide (3.4 g, 93%) as a
white solid. MS (ES+) C.sub.7H.sub.8N.sub.2O.sub.4S requires: 216.
found: 217 [M+H].sup.+.
Synthesis of 6-nitro-3,4-dihydro-1H-benzo[d][1,2]thiazine
2,2-dioxide
##STR00366##
To a mixture of (4-nitrophenyl)methanesulfonamide (864 mg, 4.0
mmol) and s-trioxane (120 mg, 4.0 mmol) in 1,2-dichoroethane (10
mL) was added trifluoromethanesulfonic acid (1.5 mL), followed by
the addition of trifluoromethanesulfonic anhydride (1.5 mL, 4.0
mmol). The reaction mixture was stirred at 35.degree. C. for 2
hours and refluxed overnight. The mixture was then diluted with
dichloromethane (100 mL) and washed with water (100 mL) and brine
(50 mL). The solvents were evaporated and the residue was purified
by Prep-HPLC to give 6-nitro-3,4-dihydro-1H-benzo[d][1,2]thiazine
2,2-dioxide (210 mg, 23%) as a gray solid. MS (ES+)
C.sub.8H.sub.8N.sub.2O.sub.4S requires: 228. found: 229
[M+H].sup.+.
Synthesis of tert-butyl
6-nitro-1,4-dihydro-3H-benzo[d][1,2]thiazine-3-carboxylate
2,2-dioxide
##STR00367##
To a solution of compound 3 (40 mg, 0.17 mmol) and
diisopropylethylamine (44 mg, 0.34 mmol) in dichloromethane (5 mL)
was added di-tert-butyl dicarbonate (57 mg, 0.26 mmol). The mixture
was then stirred at RT for 1 h, and then diluted with
dichloromethane (100 mL). The organic layer was washed with water
(100 mL) and brine (50 mL), concentrated and dried to give
tert-butyl
6-nitro-1,4-dihydro-3H-benzo[d][1,2]thiazine-3-carboxylate
2,2-dioxide (crude, 60 mg) as a yellow solid. MS (ES+)
C.sub.13H.sub.16N.sub.2O.sub.6S requires: 328. found: 351
[M+Na].sup.+.
Synthesis of tert-butyl
6-Amino-1,4-dihydro-3H-benzo[d][1,2]thiazine-3-carboxylate
2,2-dioxide
##STR00368##
A suspension of compound 4 (220 mg, 0.738 mmol) and Pd/C (50 mg) in
methanol (30 mL) was stirred at 80.degree. C. under 1 atm hydrogen
overnight. The reaction mixture was cooled to RT, and filtered
through a pad of celite. The filtrate was concentrated to afford
the tert-butyl
6-Amino-1,4-dihydro-3H-benzo[d][1,2]thiazine-3-carboxylate
2,2-dioxide (160 mg, 79.2%) as a white solid. MS (ES+)
C.sub.26H.sub.34N.sub.4O.sub.10S.sub.2 requires: 626. found: 627
[M+H].sup.+.
Synthesis of 5-bromo-2-chloropyrimidin-4-ol
##STR00369##
To a solution of 5-bromo-2,4-dichloropyrimidine (10 g, 44.2 mmol)
in THF (135 mL) was added sodium hydroxide solution (3 M, 45 mL),
and the mixture was stirred overnight at RT. The solvent was
evaporated, and the residue was diluted with water (100 mL). The
aqueous solution was cooled to 0.degree. C., brought to pH 2-3 with
1 N HCl and then extracted with methanol/dichloromethane (5%,
5.times.100 mL). The organic layers were separated, combined, dried
over sodium sulfate, filtered and concentrated to give
5-bromo-2-chloropyrimidin-4-ol (6.5 g, 71%) as a yellow solid. MS
(ES+) C.sub.4H.sub.2BrClN.sub.2O requires: 208, 210. found: 209,
211 [M+H].sup.+.
Synthesis of tert-butyl
4-(5-bromo-4-hydroxypyrimidin-2-yl)piperazine-1-carboxylate
##STR00370##
A solution of 5-bromo-2-chloropyrimidin-4-ol (6.5 g, 31 mmol),
tert-butyl piperazine-1-carboxylate (6.5 g, 35 mmol) in propan-2-ol
(200 mL) and diisopropylethylamine (10 mL) was stirred at
80.degree. C. overnight. The mixture was cooled to RT and then
concentrated. The residue was purified by silica gel chromatography
eluting with ethyl acetate:petroleum ether (100:1) to give
tert-butyl
4-(5-bromo-4-hydroxypyrimidin-2-yl)piperazine-1-carboxylate (9 g,
80%) as a yellow solid. MS (ES+) C.sub.13H.sub.19BrN.sub.4O.sub.3
requires: 358, 360. found: 303, 305 [M+H-56].sup.+.
Synthesis of 5-benzyl-2-(piperazin-1-yl)pyrimidin-4-ol
##STR00371##
To a solution of tert-butyl
4-(5-bromo-4-hydroxypyrimidin-2-yl)piperazine-1-carboxylate (20 g,
56 mmol) and Pd(Amphos)Cl.sub.2 (4.0 g 5.6 mmol) in THF (200 mL,
dry) was added benzylzinc(II) bromide (168 mL, 168 mmol) under
argon and the mixture was stirred at 60.degree. C. overnight. The
reaction mixture was diluted with ethyl acetate (1.5 L), filtered
and the filtrate was concentrated under reduced pressure. The
residue was purified by silica gel chromatography (petroleum
ether/ethyl acetate=1:1 to 100% ethyl acetate) to give
5-benzyl-2-(piperazin-1-yl)pyrimidin-4-ol (8.2 g, 54%) as a pale
yellow solid. MS (ES+) C.sub.15H.sub.18N.sub.4O requires: 270.
found: 271 [M+H].sup.+.
Synthesis of tert-butyl
4-(5-benzyl-4-hydroxypyrimidin-2-yl)piperazine-1-carboxylate
##STR00372##
To a solution of 5-benzyl-2-(piperazin-1-yl)pyrimidin-4-ol (8.0 g,
29.5 mmol) and triethylamine (8.9 g, 88.5 mmol) in THF (90 mL) was
added di-tert-butyl dicarbonate (7.7 g, 35.4 mmol). The mixture was
stirred at RT overnight, and then diluted with ethyl acetate (300
mL). The organic phase was washed with water (300 mL) and brine
(150 mL), dried over sodium sulfate, and evaporated under reduced
pressure. The residue was purified by silica gel chromatography
(petroleum ether/ethyl acetate=2:1 to 1:2) to give tert-butyl
4-(5-benzyl-4-hydroxypyrimidin-2-yl)piperazine-1-carboxylate (4.5
g, 41%) as a pale yellow solid. MS (ES+)
C.sub.20H.sub.26N.sub.4O.sub.3 requires: 370. found: 371
[M+H].sup.+.
Synthesis of tert-butyl
4-(5-benzyl-4-(trifluoromethylsulfonyloxy)pyrimidin-2-yl)piperazine-1-car-
boxylate
##STR00373##
To a solution of tert-butyl
4-(5-benzyl-4-hydroxypyrimidin-2-yl)piperazine-1-carboxylate (4.3
g, 11.5 mmol), triethylamine (3.2 mL, 23 mmol) and
N,N-dimethylpyridin-4-amine (183 mg, 1.15 mmol) in dichloromethane
(40 mL) was added trifluoromethanesulfonic anhydride (2.3 mL, 13.9
mmol). The reaction mixture was stirred at RT for 1 h, and diluted
with dichloromethane (200 mL). The organic phase was washed with
water (200 mL) and brine (100 mL), dried over sodium sulfate,
filtered and concentrated. The residue was purified by silica gel
chromatography (petroleum ether/ethyl acetate=10:1 to 5:1) to give
tert-butyl
4-(5-benzyl-4-(trifluoromethylsulfonyloxy)pyrimidin-2-yl)piperazine-1-car-
boxylate (2.4 g, 41%) as a yellow oil. MS (ES+)
C.sub.21H.sub.25F.sub.3N.sub.4O.sub.5S requires: 502. found: 525
[M+Na].sup.+.
Synthesis of tert-butyl
4-(5-benzyl-4-methylpyrimidin-2-yl)piperazine-1-carboxylate
##STR00374##
To a mixture of tert-butyl
4-(5-benzyl-4-(trifluoromethylsulfonyloxy)pyrimidin-2-yl)piperazine-1-car-
boxylate (280 mg, 0.56 mmol) and iron (III) acetylacetonate (21 mg,
0.06 mmol) in THF (5 mL, dry) and 1-methyl-2-pyrrolidinone (1 mL)
was added methylmagnesium chloride (1.68 mL, 1.68 mmol). The
reaction mixture was stirred at RT for 1 h, and diluted with ethyl
acetate (100 mL) and sat. aq. NH.sub.4Cl (100 mL). The organic
layer was washed with brine (100 mL), dried over sodium sulfate,
filtered and evaporated to give tert-butyl
4-(5-benzyl-4-methylpyrimidin-2-yl)piperazine-1-carboxylate (crude,
250 mg) as a yellow solid. MS (ES+) C.sub.21H.sub.28N.sub.4O.sub.2
requires: 368. found: 369 [M+H].sup.+.
Synthesis of 5-benzyl-4-methyl-2-(piperazin-1-yl)pyrimidine
##STR00375##
To a solution of tert-butyl
4-(5-benzyl-4-methylpyrimidin-2-yl)piperazine-1-carboxylate (250
mg, 0.68 mmol) in dioxane (2 mL) was added 4 M HCl-dioxane (2 mL).
The reaction mixture was stirred at RT for 2 h and then
concentrated to afford crude
5-benzyl-4-methyl-2-(piperazin-1-yl)pyrimidine. MS (ES+)
C.sub.16H.sub.20N.sub.4 requires: 268. found: 269 [M+H].sup.+.
Synthesis of tert-butyl
4-(5-benzylpyrimidin-2-yl)-3-(hydroxymethyl)piperazine-1-carboxylate
##STR00376##
A solution of 5-benzyl-2-chloropyrimidine (944 mg, 4.626 mmol),
tert-butyl 3-(hydroxymethyl)piperazine-1-carboxylate (1.0 g, 6.939
mmol) and diisopropylethylamine (1.8 g, 13.878 mmol) in dioxane
(100 mL) was stirred at 110.degree. C. for two days. The reaction
mixture was cooled to room temperature, concentrated and directly
purified by silica gel chromatography eluting with petroleum
ether:ethyl acetate=1:1, to afford tert-butyl
4-(5-benzylpyrimidin-2-yl)-3-(hydroxymethyl)piperazine-1-carboxylate
(400 mg, 15%) as a white solid. MS (ES+)
C.sub.21H.sub.28N.sub.4O.sub.3 requires: 384. found: 385
[M+H].sup.+.
Synthesis of (1-(5-benzylpyrimidin-2-yl)piperazin-2-yl)methanol
##STR00377##
A solution of tert-butyl
4-(5-benzylpyrimidin-2-yl)-3-(hydroxymethyl)piperazine-1-carboxylate
(400 mg, 1.042 mmol) in 4 M HCl/dioxane (20 mL) was stirred at room
temperature for 2 h. The reaction mixture was concentrated under
reduced pressure to afford
(1-(5-benzylpyrimidin-2-yl)piperazin-2-yl)methanol HCl salt (294
mg, 100%). MS (ES+) C.sub.16H.sub.20N.sub.4O requires: 284. found:
285 [M+H].sup.+.
Synthesis of benzyl
4-(5-benzoylpyrimidin-2-yl)piperazine-1-carboxylate
##STR00378##
To a solution of tert-butyl
4-(5-benzoylpyrimidin-2-yl)piperazine-1-carboxylate (957 mg, 2.6
mmol) in dioxane (20 mL) was added 4 M HCl-dioxane (20 mL). The
reaction mixture was stirred at room temperature overnight and
concentrated to afford crude
phenyl(2-(piperazin-1-yl)pyrimidin-5-yl)methanone which was used in
the next step directly.
To a solution of phenyl(2-(piperazin-1-yl)pyrimidin-5-yl)methanone
(crude, 2.6 mmol assumed) and triethylamine (780 mg, 7.8 mmol) in
dichloromethane (10 mL) was added benzyl chloroformate (663 mg, 3.9
mmol). The reaction mixture was stirred at room temperature for 2
hours, and diluted with dichloromethane (100 mL). The organic phase
was washed with water (100 mL) and brine (50 mL), dried over sodium
sulfate, filtered and concentrated. The residue was purified by
silica gel chromatography (etroleum ether/ethyl acetate=4:1-2:1) to
give benzyl 4-(5-benzoylpyrimidin-2-yl)piperazine-1-carboxylate
(600 mg, 57%) as a yellow solid. MS (ES+)
C.sub.23H.sub.22N.sub.4O.sub.3 requires: 402. found: 403
[M+H].sup.+.
Synthesis of benzyl
4-(5-(difluoro(phenyl)methyl)pyrimidin-2-yl)piperazine-1-carboxylate
##STR00379##
Benzyl 4-(5-benzoylpyrimidin-2-yl)piperazine-1-carboxylate (360 mg,
0.37 mmol) in diethylaminosulfur trifluoride (2 mL) was stirred at
90.degree. C. overnight. The reaction mixture was diluted with
ethyl acetate (100 mL), washed with sat. aq. sodium bicarbonate
(100 mL) and brine (100 mL), and evaporated in vacuum. The residue
was purified by silica gel chromatography (petroleum ether/ethyl
acetate=10:1) to give benzyl
4-(5-(difluoro(phenyl)methyl)pyrimidin-2-yl)piperazine-1-carboxylate
(180 mg, 47%) as a brown solid. MS (ES+)
C.sub.23H.sub.22F.sub.2N.sub.4O.sub.2 requires: 424. found: 425
[M+H].sup.+.
Synthesis of
5-(difluoro(phenyl)methyl)-2-(piperazin-1-yl)pyrimidine
##STR00380##
To a solution of
4-(5-(difluoro(phenyl)methyl)pyrimidin-2-yl)piperazine-1-carboxylate
(180 mg, 0.42 mmol) in chloroform (5.0 mL) was added
iodotrimethylsilane (0.8 mL) and the mixture was stirred at room
temperature for 0.5 h. The reaction was quenched by methanol (1.0
mL), followed by the addition of 2 N HCl/dioxane (2.0 mL) and
evaporated to dryness. The residue was dissolved in methanol (1.5
mL) and dropwise added into acetone (100 mL). The solid was
collected by filtration and washed with acetone (10 mL) to afford
crude 5-(difluoro(phenyl)methyl)-2-(piperazin-1-yl)pyrimidine (88
mg) as a yellow solid. MS (ES+) C.sub.15H.sub.16F.sub.2N.sub.4
requires: 290. found: 291 [M+H].sup.+.
Synthesis of
(S)-1-(5-methyl-4-nitro-1H-pyrazol-1-yl)propan-2-ol
##STR00381##
To a solution of 5-methyl-4-nitro-1H-pyrazole (2.5 g, 19.7 mmol) in
acetonitrile (100 mL) was added (S)-2-methyloxirane (1.37 g, 23.6
mmol) and cesium carbonate (19.3 g, 59.1 mmol). The mixture was
stirred at 30.degree. C. for 7 days, then cooled to room
temperature and filtered. The filtrate was concentrated under
reduced pressure, and the residue was purified by Prep-HPLC and
then chiral-HPLC to give
(S)-1-(5-methyl-4-nitro-1H-pyrazol-1-yl)propan-2-ol (0.33 g) as a
yellowish solid. MS (ES+) C.sub.7H.sub.11N.sub.3O.sub.3 requires:
185. found: 186 [M+H].sup.+.
Synthesis of
(S)-1-(4-amino-5-methyl-1H-pyrazol-1-yl)propan-2-ol
##STR00382##
To a solution of
(S)-1-(5-methyl-4-nitro-1H-pyrazol-1-yl)propan-2-ol (100 mg, 0.54
mmol) in methanol (20 mL) was added 10% Pd/C (20 mg). The reaction
mixture was stirred under 1 atm H.sub.2 atmosphere at room
temperature overnight, and filtered through a pad of Celite. The
filtrate was concentrated to give
(S)-1-(4-amino-5-methyl-1H-pyrazol-1-yl)propan-2-ol (96 mg, crude)
as a yellowish oil. MS (ES+) C.sub.7H.sub.13N.sub.3O requires: 155.
found: 156 [M+H].sup.+.
Synthesis of tert-butyl
4-(5-(1-phenylvinyl)pyrimidin-2-yl)piperazine-1-carboxylate
##STR00383##
To a solution of Ph.sub.3PCH.sub.3Br (1.69 g, 5.45 mmol) in dry THF
(20 mL) was added n-butyl lithium (2.06 mL, 2.5 M in hexane) at
room temperature. The mixture was stirred for 1.5 h, and the yellow
solution was directly used in the next reaction.
The above solution was added to a solution of tert-butyl
4-(5-benzoylpyrimidin-2-yl)piperazine-1-carboxylate (1 g, 2.7 mmol)
in dry THF (20 mL) at 0.degree. C., and the reaction solution was
stirred at room temperature for 2 h. The reaction was quenched by
water (20 mL) and extracted with ethyl acetate (200 mL). The
organic layer was separated, dried over sodium sulfate, filtered
and concentrated. The residue was purified by silica gel
chromatography eluting with petroleum ether:ethyl acetate 10:1 to
give tert-butyl
4-(5-(1-phenylvinyl)pyrimidin-2-yl)piperazine-1-carboxylate (890
mg, 89%) as a white solid. MS (ES+) C.sub.21H.sub.26N.sub.4O.sub.2
requires: 366. found: 367 [M+H].sup.+.
Synthesis of tert-butyl
4-(5-(1-phenylcyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate
##STR00384##
To a solution of tert-butyl
4-(5-(1-phenylvinyl)pyrimidin-2-yl)piperazine-1-carboxylate (366
mg, 1.0 mmol) in THF (20 mL) was added trimethylsulfoxonium iodide
(1.1 g 5.0 mmol) in dimethyl sulfoxide (7 mL). The reaction mixture
was stirred at 80.degree. C. for 4 days, and diluted with ethyl
acetate (200 mL). The organic phase was washed with water (200 mL)
and brine (100 mL), dried over sodium sulfate, and concentrated.
The residue was purified by silica gel chromatography (petroleum
ether:ethyl acetate=20:1) to give tert-butyl
4-(5-(1-phenylcyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate
(40 mg, 10%) as a yellow solid. MS (ES+)
C.sub.22H.sub.28N.sub.4O.sub.2 requires: 380. found: 381
[M+H].sup.+.
Synthesis of
5-(1-phenylcyclopropyl)-2-(piperazin-1-yl)pyrimidine
##STR00385##
To a solution of tert-butyl
4-(5-(1-phenylcyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate
(30 mg, 0.08 mmol) in dioxane (2 mL) was added 4 M HCl-dioxane (2
mL). The reaction mixture was stirred at room temperature overnight
and then concentrated to afford crude
5-(1-phenylcyclopropyl)-2-(piperazin-1-yl)pyrimidin. MS (ES+)
C.sub.17H.sub.20N.sub.4 requires: 280. found: 281 [M+H].sup.+.
Synthesis of (S,Z)-tert-butyl
4-(5-((tert-butylsulfinylimino)(4-fluorophenyl)methyl)pyrimidin-2-yl)pipe-
razine-1-carboxylate
##STR00386##
To a solution of tert-butyl
4-(5-(4-fluorobenzoyl)pyrimidin-2-yl)piperazine-1-carboxylate (4.0
g, 10.4 mmol), (S)-2-methylpropane-2-sulfinamide (2.5 g, 20.7 mmol)
in THF (60 mL) was added titanium ethoxide (20 mL) at room
temperature. The resultant solution was heated at 70.degree. C.
overnight. After that, the reaction was cooled to room temperature,
diluted with ethyl acetate (200 mL) and saturated aqueous sodium
bicarbonate (500 mL). The mixture was filtered through Celite. The
filtrate was separated. Aqueous layer was extracted with ethyl
acetate (200 mL.times.3). Then organic layers were combined, dried
over sodium sulfate, filtered and concentrated in vacuo. The
residue was purified by silica gel column (petroleum ether:ethyl
acetate 2:1) to afford the title compound (3.3 g, yield 66%) as a
yellow solid. MS (ES+) requires: 489. found 490 [M+H].sup.+;
purity: 95% (UV at 254 nm).
Synthesis of tert-butyl
4-(5-(1-((S)-1,1-dimethylethylsulfinamido)-1-(4-fluorophenyl)ethyl)pyrimi-
din-2-yl)piperazine-1-carboxylate
##STR00387##
To a solution of (S,Z)-tert-butyl
4-(5-((tert-butylsulfinylimino)(4-fluorophenyl)methyl)pyrimidin-2-yl)pipe-
razine-1-carboxylate (3.0 g, 6.1 mmol) in THF (20 mL) was dropwise
added methyl magnesium bromide (2.1 mL, 6.1 mmol, 3 M in ethyl
ether) slowly at -60.degree. C. under nitrogen. After addition, the
mixture was allowed to warm to room temperature and stirred for 2
h. Then, the reaction was quenched by methanol (20 mL) and
saturated aqueous ammonium chloride (50 mL), and extracted with
ethyl acetate (50 mL.times.3). The combined organic layers were
dried over sodium sulfate, filtered and concentrated in vacuo. The
residue was purified by silica gel column
(dichloromethane:methanol=40:1) to give the title compound 3 (2.5
g, yield 77%) as a yellow solid. MS (ES+) requires: 505. found 506
[M+Na].sup.+; purity: 100% (UV at 254 nm).
Preparation of
(S)-1-(4-fluorophenyl)-1-(2-(piperazin-1-yl)pyrimidin-5-yl)ethanamine
##STR00388##
To a solution of tert-butyl
4-(5-(1-((S)-1,1-dimethylethylsulfinamido)-1-(4-fluorophenyl)ethyl)pyrimi-
din-2-yl)piperazine-1-carboxylate (2.5 g, 5.0 mmol) in dioxane (5.0
mL) was dropwise added HCl (4M in dioxane, 10 mL). The reaction
solution was stirred at room temperature overnight. Then the
solution was concentrated under vacuo to give the title compound
(1.5 g, crude yield 100%) as a yellow solid. MS (ES+)
C.sub.16H.sub.20FN.sub.5 requires: 301, found 302 [M+H].sup.+;
purity: 100% (UV at 254 nm). The above racemate (1.5 g) was
separated by Chiral-HPLC to afford the desired single enantiomer
(600 mg, 40%) as a yellow solid. MS (ES+) C.sub.16H.sub.20FN.sub.5
requires: 301. found: 302 [M+H].sup.+.
The synthetic protocol that can be used to prepare the compounds
disclosed herein is shown below. The NMR and LC MS data obtained
for compounds disclosed herein are also shown below.
TABLE-US-00002 Compound Synthetic LC/MS Number Protocol .sup.1H NMR
M + 1 4 3 This spectrum contains some rotamers in the aromatic
region - 1H 416 NMR (400 MHz, DMSO-d6) .delta. 12.54 (s, 1H), 9.52
(d, J = 61.6 Hz, 2H), 8.30 (s, 2H), 8.14 (s, 1H), 7.82 (s, 1H),
7.55 (d, J = 21.1 Hz, 1H), 7.34-7.11 (m, 5H), 3.80 (q, J = 9.2, 7.5
Hz, 10H). 5 3 416 6 2 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 9.66, 9.53 (s, s, 1H), 8.40 (d, 2H, J = 4.5 Hz), 8.31, 8.17 (s,
s, 1H), 7.84 (s, 1H), 7.51, 7.50 (s, s, 1H), 6.68 (t, 1H, J = 5.0
Hz), 4.11-4.07 (m, 2H), 3.86-3.66 (m, 10H), 3.39-3.34 (m, 1H),
2.67-2.64 (m, 3H), 2.38-2.34 (br, 1H). 7 3 1H NMR (400 MHz,
DMSO-d6) .delta. 9.75-9.62 (m, 1H), 8.31 (s, 426 2H), 8.26 (s, 1H),
7.75-7.66 (m, 2H), 7.37-7.13 (m, 7H), 7.04-6.93 (m, 1H), 3.93-3.68
(m, 10H). 9 5 426 10 3 This spectrum contains some rotamers in the
aromatic region: 1H 429 NMR (400 MHz, DMSO-d6) .delta. 9.62, 9.48
(s, 1H), 8.31, 8.26 (s, 2H), 8.14 (s, 1H), 7.82, 7.78 (s, 1H),
7.47, 7.42 (s, 1H), 7.34-7.11 (m, 5H), 3.87-3.71 (m, 10H). 11 3
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.42 (s, 1H), 8.31 (m, 3H),
430 7.34-7.15 (m, 5H), 6.71 (s, 1H), 3.81 (m, 10H), 3.41-3.22 (m,
2H), 2.40 (m, 3H). 12 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.38
(s, 1H), 8.31 (d, J = 1.0 Hz, 440 2H), 8.22 (s, 1H), 7.34-7.14 (m,
5H), 7.06 (s, 1H), 6.91 (t, J = 7.9 Hz, 1H), 6.82-6.76 (m, 1H),
6.27-6.19 (m, 1H), 5.12 (s, 2H), 3.89-3.72 (m, 10H). 13 3 .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 9.60 (s, 1H), 8.31 (s, 2H), 441 8.25
(s, 1H), 7.99 (d, J = 2.1 Hz, 1H), 7.58 (d, J = 2.4 Hz, 1H), 7.44
(t, J = 2.4 Hz, 1H), 7.33-7.14 (m, 5H), 5.37 (s, 2H), 3.80 (m,
10H). 14 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.62 (s, 1H),
8.31 (s, 2H), 454 8.25 (s, 1H), 7.78 (s, 1H), 7.45 (d, J = 8.1 Hz,
1H), 7.35-7.14 (m, 6H), 6.96 (d, J = 7.6 Hz, 1H), 3.87-3.74 (m,
10H), 3.70 (s, 2H). 15 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.66 (s, 1H), 8.31 (s, 2H), 455 8.25 (s, 1H), 7.83 (s, 1H), 7.47
(d, J = 8.1 Hz, 1H), 7.34-7.13 (m, 6H), 6.92 (d, J = 7.5 Hz, 1H),
5.17 (t, J = 5.7 Hz, 1H), 4.47 (d, J = 5.7 Hz, 2H), 3.91-3.70 (m,
10H). 16 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.64 (s, 1H),
8.31 (s, 2H), 455 8.25 (s, 1H), 7.64 (dd, J = 8.6, 2.4 Hz, 2H),
7.35-7.14 (m, 7H), 4.43 (s, 2H), 3.90-3.72 (m, 10H). 17 3
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 10.05 (s, 1H), 8.36
(s, 1H), 8.32 (s, 2H), 7.98 (d, 1H, J = 5.6 Hz), 7.31-7.18 (m, 7H),
3.86-3.76 (m, 13H). 19 1 .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 9.67, 9.53 (s, s, 1H), 8.29-8.16 (m, 3H), 7.83 (s, 1H),
7.51-7.48 (m, 1H), 4.21-4.05 (m, 2H), 3.83-3.68 (m, 11H), 3.51-3.42
(m, 1H), 2.82-2.63 (m, 3H), 2.45-2.39 (m, 1H), 1.80-1.76 (m, 1H),
0.89-0.86 (m, 2H), 0.66-0.63 (m, 2H). 22 3 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.90 (s, 1H), 8.53 (br.s, 1H), 467 8.31 (s, 2H),
8.29 (s, 1H), 7.82 (d, J = 8.0 Hz, 1H), 7.61 (dd, J = 7.8, 1.7 Hz,
1H), 7.45 (t, J = 7.9 Hz, 1H), 7.29 (t, J = 7.4 Hz, 2H), 7.25-7.14
(m, 3H), 3.94-3.72 (m, 14H), 2.57 (s, 3H). 23 3 .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.99 (s, 1H), 8.39 (s, 1H), 468 8.32 (d,
J = 1.4 Hz, 2H), 7.91 (s, 1H), 7.70 (dd, J = 8.0, 2.2 Hz, 1H),
7.56-7.48 (m, 1H), 7.44-7.12 (m, 6H), 3.94-3.69 (m, 10H). 24 3
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.91 (s, 1H), 8.31 (s, 3H),
468 7.88-7.72 (m, 5H), 7.35-7.12 (m, 6H), 3.91-3.73 (m, 10H). 25 3
468 26 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.64 (s, 1H), 8.31
(s, 2H), 469 8.25 (s, 1H), 7.90 (s, 1H), 7.41 (s, 1H), 7.32-7.25
(m, 2H), 7.25-7.15 (m, 4H), 6.95 (d, J = 7.5 Hz, 1H), 5.14 (d, J =
4.0 Hz, 1H), 4.73-4.64 (m, 1H), 3.82 (d, J = 21.7 Hz, 10H), 1.31
(d, J = 6.4 Hz, 3H). 27 3 .sup.1H-NMR (500 MHz, DMSO-d.sub.6)
.delta. 9.72, 9.59 (br. s., br. s., 1H), 8.32-8.30 (m, 2H), 8.17
(s, 1H), 7.99-7.96 (m, 1H), 7.61-7.55 (m, 1H), 7.31-7.18 (m, 5H),
5.22-5.13 (m, 1H), 4.34-3.69 (m, 15H). 28 3 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. 9.72, 9.58 (s, s, 1H), 8.32-8.29 (m, 2H),
8.17 (s, 1H), 8.00, 7.94 (s, s, 1H), 7.67, 7.61 (s, s, 1H),
7.35-7.15 (m, 5H), 5.66-5.44 (m, 1H), 4.96-4.79 (m, 4H), 3.84-3.78
(m, 10H). 29 4 .sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm 9.70 (s,
1H), 8.39 (s, 2H), 472 8.27 (s, 1H), 7.72-7.70 (m, 2H), 7.48-7.44
(m, 2H), 7.32-7.29 (m, 2H), 7.13-7.08 (m, 2H), 7.00 (t, 1H, J = 7.6
Hz), 3.85-3.72 (m, 8H), 2.55 (br, 2H), 1.72 (s, 3H). 30 3 This
spectrum contains some rotamers in the aromatic region: 1H 472 NMR
(400 MHz, DMSO-d6) .delta. 9.61, 9.48 (s, 1H), 8.30, 8.23 (s, 2H),
8.14 (s, 1H), 7.87, 7.85 (s, 1H), 7.47, 7.45 (s, 1H), 7.32-7.13 (m,
5H), 4.13 (t, J = 6.1 Hz, 2H), 3.79 (m, 10H), 2.82 (t, J = 6.0 Hz,
2H), 2.27 (s, 3H). 31 3 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 9.63, 9.48 (s, s, 1H), 8.32 (s, 2H), 8.16 (s, 1H), 7.86 (s,
1H), 7.49 (s, 1H), 7.29-7.19 (m, 5H), 4.25-4.19 (m, 2H), 3.94-3.79
(m, 10H), 3.66 (s, 2H), 3.25 (s, 3H). 32 3 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 8.33-8.27 (m, 3H), 8.15, 8.05 (s, s, 1H),
7.90, 7.88 (s, s, 1H), 7.49 (s, 1H), 7.32-7.20 (m, 5H), 4.03-3.90
(m, 3H), 3.80 (m, 11H), 1.04 (d, 3H, J = 6.0 Hz). 33 3 .sup.1H-NMR
(500 MHz, DMSO-d.sub.6) .delta. ppm 8.34-8.26 (m, 3H), 8.15 (s,
1H), 7.90, 7.87 (s, s, 1H), 7.49 (s, 1H), 7.31-7.20 (m, 5H),
4.01-3.94 (m, 3H), 3.82-3.74 (m, 11H), 1.03 (d, 3H, J = 6.0 Hz). 36
4 .sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm 9.64, 9.50 (s, 1H),
8.38 (s, 477 2H), 8.27, 8.14 (s, 1H), 7.82, 7.79 (s, 1H), 7.47-7.43
(m, 3H), 7.11-7.07 (m, 2H), 3.82-3.77 (m, 11H), 2.49 (s, 2H), 1.71
(s, 3H). 37 2 476 38 4 .sup.1H-NMR (400 MHz, CDCl3) .delta. ppm
8.35 (s, 2H), 8.21 (br. s., 1H), 477 7.80, 7.22 (br. s., br. s.,
1H), 7.61-7.55 (m, 1H), 7.43-7.38 (m, 2H), 7.04-7.00 (m, 2H), 3.90
(br. s., 11H), 2.30 (br. s., 1H), 1.92 (s, 3H). 39 4 .sup.1H-NMR
(400 MHz, CDCl3) .delta. ppm 8.35 (s., 2H), 8.20 (br. s., 1H), 477
7.77, 7.32 (br. s., br. s., 1H), 7.61-7.56 (m, 1H), 7.42-7.38 (m,
2H), 7.05-7.00 (m, 2H), 3.89 (br. s., 11H), 2.32 (br. s., 1H), 1.92
(s., 3H). 41 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.55 (s, 1H),
8.31, 8.30 (s, 2H), 480 8.24 (s, 1H), 7.42 (s, 1H), 7.37 (dd, J =
8.2, 2.3 Hz, 1H), 7.34-7.25 (m, 2H), 7.25-7.15 (m, 3H), 6.99 (d, J
= 8.4 Hz, 1H), 3.89-3.71 (m, 12H), 2.96 (t, J = 5.9 Hz, 2H), 2.64
(t, J = 5.9 Hz, 2H). 42 3 480 43 3 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.73 (s, 1H), 8.61, 8.56 (s, 1H), 481 8.32, 8.30 (s, 2H),
8.27 (s, 1H), 7.86-7.73 (m, 1H), 7.35-7.14 (m, 5H), 3.97-3.65 (m,
12H), 3.00 (t, J = 6.0 Hz, 1H), 2.94-2.79 (m, 2H), 2.70 (t, J = 5.8
Hz, 1H). 44 1 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.56 (s, 1H),
8.52-8.42 (m, 481 1H), 8.35 (s, 2H), 8.24 (s, 1H), 7.76-7.67 (m,
2H), 7.48-7.34 (m, 2H), 7.31 (d, J = 7.7 Hz, 1H), 7.21 (dd, J =
7.5, 4.7 Hz, 1H), 7.01 (d, J = 8.3 Hz, 1H), 3.94 (s, 2H), 3.88 (s,
2H), 3.80 (br.s., 8H), 3.00 (t, J = 6.0 Hz, 2H), 2.67 (t, J = 5.9
Hz, 2H). 45 1 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.55
(br, 1H), 8.52 (s, 1H), 8.42 (d, 1H, J = 4.5 Hz), 8.35 (s, 2H),
8.24 (s, 1H), 7.64 (d, 1H, J = 8.0 Hz), 7.41 (s, 1H), 7.37 (d, 1H,
J = 8.0 Hz), 7.31 (dd, 1H, J = 8.0, 4.7 Hz), 6.99 (d, 1H, J = 8.0
Hz), 3.83 (br. s., 12H), 2.95 (d, 2H, J = 6.0 Hz), 2.63 (t, 2H, J =
6.0 Hz). 46 5 .sup.1H NMR (400 MHz, Methanol-d4) .delta. 8.22 (s,
3H), 7.61-7.48 (m, 482 2H), 7.39-7.29 (m, 2H), 7.22 (d, J = 8.4 Hz,
1H), 7.09 (tt, J = 7.3, 1.1 Hz, 1H), 7.00-6.92 (m, 2H), 4.36 (s,
2H), 3.98-3.84 (m, 8H), 3.51 (t, J = 6.4 Hz, 2H), 3.09 (t, J = 6.4
Hz, 2H). 47 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.87 (s, 1H),
9.65 (s, 1H), 482 8.31 (s, 2H), 8.24 (s, 1H), 7.38-7.01 (m, 9H),
3.82 (m, 10H), 2.03 (s, 3H). 48 3 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.66 (s, 1H), 8.31 (s, 2H), 482 8.25 (d, J = 2.0 Hz, 2H),
7.85 (s, 1H), 7.43 (s, 1H), 7.34-7.14 (m, 6H), 6.98 (d, J = 7.6 Hz,
1H), 3.87-3.75 (m, 10H), 3.65 (q, J = 6.5 Hz, 1H), 2.18 (s, 3H),
1.28 (d, J = 6.6 Hz, 3H). 49 3 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.65 (s, 1H), 8.31 (s, 2H), 482 8.25 (s, 1H), 7.86 (s, 1H),
7.45 (d, J = 8.4 Hz, 1H), 7.33-7.14 (m, 6H), 6.89 (d, J = 7.5 Hz,
1H), 3.93-3.70 (m, 10H), 3.34 (s, 2H), 2.15 (s, 6H). 50 3 .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 9.62 (s, 1H), 8.31 (s, 2H), 483 8.24
(s, 1H), 7.36 (s, 1H), 7.28 (t, J = 7.4 Hz, 2H), 7.21 (m, 4H), 7.09
(d, J = 7.6 Hz, 1H), 4.97 (s, 1H), 3.82 (m, 10H), 3.65-3.53 (m,
3H), 3.19-3.07 (m, 3H), 1.42 (s, 6H). 51 3 This spectrum contains
some rotamers in the aromatic region: 1H 484 NMR (400 MHz, DMSO-d6)
.delta. 9.61, 9.47 (s, 1H), 8.30, 8.25 (s, 2H), 8.13 (s, 1H), 7.81
(s, 1H), 7.45 (d, J = 11.5 Hz, 1H), 7.32-7.13 (m, 5H), 4.25 (dd, J
= 23.2, 7.4 Hz, 2H), 3.88-3.70 (m, 10H), 3.53-3.42 (m, 2H),
3.05-2.85 (m, 2H) - one peak is obscured partially by the water
signal. 52 3 This spectrum contains some rotamers in the aromatic
region: 1H 484 NMR (400 MHz, DMSO-d6) .delta. 9.68-9.45 (m, 1H),
8.35-8.23 (m, 2H), 8.14 (d, J = 4.5 Hz, 1H), 7.89 (s, 1H), 7.48 (d,
J = 18.8 Hz, 1H), 7.31-7.14 (m, 5H), 4.91-4.77 (mz, 1H), 3.90-3.67
(m, 10H), 3.20-3.10 (m, 1H), 3.07-2.80 (m, 3H), 2.76-2.58 (m, 0H),
2.40-2.22 (m, 1H), 2.17 (dd, J = 13.8, 7.2 Hz, 1H), 1.98 (s, 1H).
53 3 This spectrum contains some rotamers in the aromatic region:
1H 484 NMR (400 MHz, DMSO-d6) .delta. 9.71-9.44 (m, 1H), 8.35-8.22
(m, 2H), 8.13 (d, J = 2.8 Hz, 1H), 7.94-7.82 (m, 1H), 7.57-7.39 (m,
1H), 7.33-7.11 (m, 5H), 4.92-4.75 (m, 1H), 3.89-3.70 (m, 10H),
3.08-2.94 (m, 1H), 2.94-2.76 (m, 2H), 2.76-2.60 (m, 1H), 2.40-2.24
(m, 1H), 2.23-2.07 (m, 1H), 2.07-1.89 (m, 1H). 54 3 485 55 3
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.51 (s, 1H), 8.32 (s, 2H),
485 7.56 (d, J = 8.5 Hz, 2H), 7.37-7.08 (m, 5H), 6.89 (d, J = 8.5
Hz, 2H), 4.92 (d, J = 4.9 Hz, 1H), 4.65 (t, J = 5.6 Hz, 1H), 3.96
(dd, J = 9.5, 4.0 Hz, 1H), 3.80 (m, 10H), 3.44 (t, J = 5.5 Hz, 2H).
56 3 .sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm 9.64 (s, 1H), 8.31
(s, 2H), 485 8.25 (s, 1H), 7.63 (d, 2H, J = 8.4 Hz), 7.31-7.17 (m,
7H), 5.15 (d, 1H, J = 4.0 Hz), 4.69 (t, 1H, J = 5.6 Hz), 4.49 (dd,
1H, J = 10 Hz, J = 5.6 Hz), 3.83-3.80 (m, 10H), 3.39 (t, 1H, J =
5.6 Hz). 57 3 .sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm 9.65 (s,
1H), 8.32 (s, 2H), 485 8.26 (s, 1H), 7.63 (d, 2H, J = 8.4 Hz),
7.31-7.17 (m, 7H), 5.15 (d, 1H, J = 4.0 Hz), 4.69 (t, 1H, J = 5.6
Hz), 4.49 (dd, 1H, J = 10 Hz, J = 5.6 Hz), 3.83-3.80 (m, 10H), 3.41
(t, 1H, J = 5.6 Hz). 58 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.33 (s, 1H), 8.31 (s, 2H), 485 8.17 (s, 1H), 7.62 (s, 1H), 7.29
(mz, 2H), 7.26-7.14 (m, 3H), 6.49 (d, J = 8.9 Hz, 1H), 6.30 (t, J =
5.8 Hz, 1H), 4.70 (t, J = 5.3 Hz, 1H), 3.78 (m, 10H), 3.55-3.46 (m,
2H) - one peak obscured by water signal. 60 3 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.61 (s, 1H), 8.31 (s, 2H), 486 8.13 (d, J = 7.8
Hz, 1H), 7.87 (d, J = 13.9 Hz, 1H), 7.45 (s, 1H), 7.30-7.15 (m,
5H), 4.22-4.13 (m, 2H), 3.81 (d, J = 15.3 Hz, 10H), 2.59 (t, J =
6.3 Hz, 2H), 2.20-2.11 (m, 6H). 61 3 This spectrum contains some
rotomers in the aromatic region: .sup.1H 486 NMR (400 MHz, DMSO-d6)
.delta. 9.61, 9.48 (s, 1H), 8.30, 8.27 (s, 2H), 8.14 (s, 1H), 7.87,
7.84 (s, 1H), 7.47, 7.45 (s, 1H), 7.31-7.25 (m, 2H), 7.24-7.15 (m,
3H), 4.20-4.02 (m, 3H), 3.79 (t, J = 11.9 Hz, 12H), 2.82 (t, J =
6.0 Hz, 2H), 2.27 (s, 3H). 62 3 .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 9.64, 9.52 (s, s, 1H), 8.32 (s, 2H), 8.16 (s, 1H),
7.94, 7.90 (s, s, 1H), 7.45 (s, 1H), 7.34-7.26 (m, 2H), 7.25-7.14
(m, 3H), 4.71, 4.69 (s, s, 1H), 3.99, 3.97 (s, s, 2H), 3.82-3.77
(m, 10H), 1.05 (s, 6H). 63 6 .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 9.67, 9.54 (s, s, 1H), 8.42 (s, 2H), 8.29, 8.16 (s, s,
1H), 7.82 (s, 1H), 7.50, 7.48 (s, s, 1H),
4.13-4.04 (m, 2H), 3.85-3.69 (m, 11H), 3.44-3.39 (m, 1H), 2.77-2.63
(m, 3H), 2.42-2.37 (m, 1H), 1.56-1.52 (m, 1H), 0.89-0.85 (m, 2H),
0.73-0.69 (m, 2H). 64 3 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 9.62, 9.49 (s, s, 1H), 8.32 (s, 2H), 8.28, 8.15 (s, s, 1H),
7.90, 7.86 (s, s, 1H), 7.49 (s, 1H), 7.29 (t, 2H, J = 7.0 Hz), 7.23
(d, 2H, J = 7.0 Hz), 7.19 (t, 1H, J = 7.0 Hz), 5.01-4.84 (m, 1H),
4.74-4.71 (m, 1H), 4.21-4.17 (m, 1H), 4.00-3.95 (m, 1H), 3.83-3.76
(m, 11H). 65 3 .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
9.63, 9.49 (s, s, 1H), 8.32 (s, 2H), 8.28, 8.15 (s, 1H), 7.90, 7.86
(s, s, 1H), 7.46 (s, 1H), 7.31-7.17 (m, 5H), 5.01-4.98 (m, 1H),
4.72 (dd, 1H, J = 11.6, 5.6 Hz), 4.18 (dd, 1H, J = 14.0, 3.6 Hz),
3.97 (dd, 1H, J = 14.4, 7.6 Hz), 3.84-3.72 (m, 11H), 3.33-3.30 (m,
2H). 66 3 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.65,
9.52 (s, s, 1H), 8.32 (s, 2H), 8.29, 8.16 (s, s, 1H), 7.91, 7.86
(s, s, 1H), 7.48 (s, 1H), 7.31-7.18 (m, 5H), 5.03-4.96 (m, 1H),
4.76-4.72 (m, 1H), 4.19 (dd, 1H, J = 13.5, 3.5 Hz), 3.98 (dd, 1H, J
= 14.0, 7.0 Hz), 3.83-3.72 (m, 11H), 3.33-3.30 (m, 2H). 67 6
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.65, 9.52 (s, s,
1H), 8.29 (s, 2H), 8.16 (s, 1H), 7.83 (s, 1H), 7.50, 7.47-7.47 (s,
s, 1H), 4.12-4.03 (m, 2H), 3.84-3.63 (m, 10H), 3.42-3.39 (m, 3H),
2.72-2.57 (m, 3H), 2.33 (t, 1H, J = 10.8 Hz), 1.44-1.39 (m, 2H),
0.67-0.61 (m, 1H), 0.40-0.38 (m, 2H), 0.03-0.00 (m, 2H). 68 3
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.20 (s, 1H), 9.63 (s, 1H),
494 8.32 (d, J = 1.0 Hz, 2H), 8.23 (s, 1H), 7.40 (s, 1H), 7.34-7.15
(m, 5H), 7.07 (s, 2H), 3.81 (m, 10H), 2.80 (t, J = 7.5 Hz, 2H),
2.42 (t, J = 7.6 Hz, 2H). 69 3 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.54 (s, 1H), 8.30 (s, 2H), 494 8.23 (s, 1H), 7.45-7.35 (m,
2H), 7.31-7.25 (m, 2H), 7.24-7.16 (m, 3H), 7.01 (d, J = 8.2 Hz,
1H), 3.79 (d, J = 6.8 Hz, 10H), 3.44 (s, 2H), 2.74 (t, J = 6.0 Hz,
2H), 2.57 (t, J = 5.9 Hz, 2H), 2.33 (s, 3H). 70 3 .sup.1H-NMR (500
MHz, DMSO-d.sub.6) .delta. ppm 9.53 (br. s., 1H), 8.25 (s, 1H),
8.18 (s, 1H), 7.40-7.37 (m, 2H), 7.31-7.28 (m, 2H), 7.21-7.15 (m,
3H), 6.99 (d, 1H, J = 8.5 Hz), 3.84-3.82 (m, 12H), 2.93 (t, 2H, J =
5.5 Hz), 2.64-2.62 (m, 2H), 2.22 (s, 3H). 72 3 .sup.1H NMR (400
MHz, DMSO-d6) .delta. 9.67 (s, 1H), 8.37-8.23 (m, 494 3H),
7.57-7.44 (m, 2H), 7.32-7.13 (m, 6H), 4.00 (s, 2H), 3.80 (d, J =
11.6 Hz, 10H), 3.17 (t, J = 5.3 Hz, 2H), 2.91-2.83 (m, 2H), 1.75
(q, J = 6.4, 5.3 Hz, 2H). 73 1 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.54 (s, 1H), 8.22 (s, 1H), 495 7.64 (s, 1H), 7.46 (s, 1H),
7.40-7.33 (m, 1H), 7.28-7.20 (m, 4H), 7.19-7.13 (m, 1H), 7.00 (d, J
= 8.2 Hz, 1H), 6.30 (s, 2H), 3.89 (s, 2H), 3.81-3.63 (m, 12H), 3.00
(t, J = 5.9 Hz, 2H), 2.67 (t, J = 5.9 Hz, 2H). 74 3 .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 9.67 (s, 1H), 8.31 (m, 3H), 496 8.25 (s,
1H), 7.68 (s, 1H), 7.49 (d, J = 8.0 Hz, 1H), 7.34-7.12 (m, 6H),
6.89 (d, J = 7.6 Hz, 1H), 4.22 (d, J = 5.9 Hz, 2H), 3.80 (d, J =
12.2 Hz, 10H), 1.87 (s, 3H). 75 2 496 76 3 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.64 (s, 1H), 8.31 (s, 2H), 496 8.24 (s, 1H), 7.62
(d, J = 2.7 Hz, 1H), 7.48 (dd, J = 8.6, 2.6 Hz, 1H), 7.33-7.25 (m,
2H), 7.25-7.13 (m, 3H), 6.97 (d, J = 8.6 Hz, 1H), 4.06-3.93 (m,
4H), 3.80 (d, J = 8.9 Hz, 10H), 3.22 (t, J = 4.4 Hz, 3H). 77 2
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.51 (s, 1H), 8.30 (s, 2H),
496 8.20 (s, 1H), 7.61-7.49 (m, 2H), 7.34-7.14 (m, 5H), 6.74 (d, J
= 8.7 Hz, 2H), 4.97-4.83 (m, 1H), 3.87-3.69 (m, 12H), 3.54-3.43 (m,
2H). 78 1 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.70 (s, 1H),
8.60-8.46 (m, 496 1H), 8.26 (s, 1H), 7.79 (d, J = 2.4 Hz, 1H), 7.64
(s, 1H), 7.33-7.11 (m, 5H), 6.31 (s, 2H), 3.90-3.57 (m, 12H), 3.00
(t, J = 6.0 Hz, 2H), 2.70 (t, J = 6.0 Hz, 2H). 79 3 .sup.1H-NMR
(400 MHz, DMSO-d6) .delta. ppm 9.74 (s, 1H), 8.32 (s, 2H), 497 8.27
(s, 1H), 7.72 (d, 2H, J = 8.4 Hz), 7.52 (d, 2H, J = 8.4 Hz),
7.32-7.17 (m, 5H), 6.26 (s, 1H), 4.75 (d, 2H, J = 6.0 Hz), 4.69 (d,
2H, J = 6.4 Hz), 3.89-3.74 (m, 10H). 80 1 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.54 (br, 1H), 8.34 (s, 2H), 8.24 (s,
1H), 7.44-7.29 (m, 3H), 7.11-7.08 (m, 2H), 7.04-6.97 (m, 2H), 3.82
(br, 13H), 2.94-2.91 (br, 2H), 2.53-2.48 (m, 2H). 81 2 .sup.1H-NMR
(500 MHz, DMSO-d.sub.6) .delta. ppm 9.32, 9.17 (br. s., br. s.,
1H), 8.32 (s, 2H), 8.16 (s, 1H), 7.35-7.18 (m, 7H), 6.55 (br, 2H),
5.30-5.26 (br, 1H), 4.68 (br. s., 1H), 3.80-3.77 (m, 11H),
2.93-2.86 (m, 2H), 1.11 (d, 3H, J = 5.5 Hz). 82 2 .sup.1H-NMR (500
MHz, DMSO-d.sub.6) .delta. ppm 9.31, 9.15 (br., s., br. s., 1H),
8.31 (s, 2H), 8.15 (s, 1H), 7.34-7.17 (m, 7H), 6.55 (br. s., 2H),
5.26-5.24 (m, 1H), 4.66 (d, 1H, J = 5.0 Hz), 3.79-3.77 (m, 11H),
2.95-2.88 (m, 2H), 1.10 (d, 3H, J = 6.0 Hz). 83 3 This spectrum
contains some rotamers in the aromatic region: .sup.1H 498 NMR (400
MHz, DMSO-d6) .delta. 9.61, 9.49 (s, 1H), 8.30, 8.27 (s, 2H), 8.14
(s, 1H), 7.82 (s, 1H), 7.49 (s, 1H), 7.23 (tt, J = 20.6, 7.2 Hz,
5H), 4.08-3.93 (m, 2H), 3.79 (m 11H), 2.90-2.70 (m, 2H), 1.83-1.69
(m, 1H), 1.40 (dd, J = 16.3, 9.3 Hz, 1H) - one peak is obscured by
DMSO. 84 3 This spectrum contains some rotamers in the aromatic
region: .sup.1H 498 NMR (400 MHz, DMSO-d6) .delta. 9.61, 9.49 (s,
1H), 8.31, 8.27 (s, 2H), 8.14 (s, 1H), 7.91, 7.87 (s, 1H), 7.47 (s,
1H), 7.35-7.11 (m, 6H), 4.06-3.91 (m, 3H), 3.81 (mz, 10H),
2.85-2.70 (m, 2H), 1.76-1.53 (m, 3H), 1.41-1.27 (m, 1H). 85 3 This
spectrum contains some rotamers in the aromatic region: .sup.1H 498
NMR (400 MHz, DMSO-d6) .delta. 9.61, 9.49 (s, 1H), 8.3, 8.27 (s,
2H), 8.14 (s, 1H), 7.91, 7.87 (s, 1H), 7.46 (d, J = 3.8 Hz, 1H),
7.32-7.25 (m, 2H), 7.25-7.15 (m, 3H), 4.05-3.92 (m, 2H), 3.81 (m,
10H), 2.85-2.71 (m, 2H), 1.77-1.50 (m, 3H), 1.41-1.26 (m, 1H) - a
peak is partially obscured by water signal. 86 3 .sup.1H-NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 9.65, 9.54 (s, s, 1H), 8.32 (s, 2H),
8.29, 8.16 (s, s, 1H), 7.89, 7.86 (s, s, 1H), 7.51, 7.47 (s, s,
1H), 7.31-7.17 (m, 5H), 4.12-4.03 (m, 2H), 3.83-3.73 (m, 10H),
3.66-3.60 (m, 2H), 3.48-3.45 (m, 2H), 2.72-2.63 (m, 1H), 1.93-1.87
(m, 1H), 1.62-1.55 (m, 1H). 87 3 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.65, 9.59 (s, s, 1H), 8.33-8.31 (m, 2H),
8.16 (s, 1H), 7.87 (s, 1H), 7.50 (s, 1H), 7.34-7.18 (m, 5H), 4.59
(d, 2H, J = 5.6 Hz), 4.35-4.22 (m, 4H), 3.85-3.75 (m, 10H), 1.13
(s, 3H). 88 3 This spectrum contains some rotamers in the aromatic
region: 1H 500 NMR (400 MHz, DMSO-d6) .delta. 9.61, 9.48 (s, 1H),
8.31, 8.27 (s, 2H), 8.14 (s, 1H), 7.83, 7.79 (s, 1H), 7.50, 7.45
(s, 1H), 7.32-7.13 (m, 5H), 4.12-4.02 (m, 2H), 3.80 (d, J = 12.9
Hz, 10H), 2.13 (s, 8H), 1.93-1.80 (m, 2H). 89 3 .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.94 (s, 1H), 8.31 (d, J = 1.8 Hz, 501
3H - actually two closely spaced singlets), 7.89-7.81 (m, 2H), 7.68
(dd, J = 11.0, 8.4 Hz, 2H), 7.35-7.14 (m, 5H), 3.88-3.75 (m, 10H),
1.62 (s, 3H), 1.59 (s, 3H). 91 1 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.66, 9.60 (s, s, 1H), 8.52 (s, 2H),
8.24, 8.17 (s, s, 1H), 7.83 (s, 1H), 7.50, 7.49 (s, s, 1H),
4.18-4.01 (m, 2H), 3.92-3.62 (m, 10H), 3.45-3.37 (m, 1H), 2.75-2.56
(m, 3H), 2.39-2.28 (m, 1H). 92 2 .sup.1H-NMR (400 MHz, DMSO-d6)
.delta. ppm 9.63 (br. s., 1H), 8.32 (s, 503 2H), 8.25 (s, 1H), 7.62
(d, 2H, J = 8.8 Hz), 7.29-7.25 (m, 4H), 7.14-7.09 (m, 2H), 5.12 (d,
1H, J = 4.0 Hz), 4.68-4.65 (m, 1H), 4.50-4.46 (m, 1H), 3.83 (s,
5H), 3.79 (s, 5H), 3.42-3.39 (m, 2H). 93 2 .sup.1H-NMR (400 MHz,
DMSO-d6) .delta. ppm 9.62 (br. s., 1H), 8.31 (s, 503 2H), 8.25 (s,
1H), 7.62 (d, 2H, J = 8.8 Hz), 7.29-7.25 (m, 4H), 7.14-7.09 (m,
2H), 5.11 (br. s., 1H), 4.66 (br. s., 1H, J = 6.0 Hz), 4.50-4.46
(m, 1H), 3.84-3.79 (m, 10H), 3.42-3.40 (m, 2H). 94 3 .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 10.21 (s, 1H), 8.36 (s, 1H), 8.32 (s,
2H), 8.02-7.93 (m, 2H), 7.89-7.81 (m, 2H), 7.34-7.14 (m, 5H), 3.85
(br.s, 10H), 3.15 (s, 3H). 95 3 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 10.05 (s, 1H), 8.60 (s, 1H), 504 8.32 (s, 2H), 8.31 (s,
1H), 7.66 (d, J = 7.6 Hz, 1H), 7.51-7.43 (m, 2H), 7.38-7.12 (m,
7H), 3.92-3.73 (m, 14H). 96 3 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 10.03 (s, 1H), 8.32 (s, 1H), 504 8.31 (s, 2H), 7.91-7.84
(m, 2H), 7.78-7.71 (m, 2H), 7.32-7.14 (m, 7H), 3.92-3.68 (m, 10H).
98 3 508 99 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.31 (s, 1H),
9.62 (s, 2H), 508 8.37 (d, J = 12.4 Hz, 3H), 7.57-7.41 (m, 3H),
7.33-7.14 (m, 5H), 4.23 (d, J = 4.5 Hz, 2H), 3.81 (m, 10H), 3.17
(dd, J = 6.8, 2.8 Hz, 2H), 1.34 (s, 6H). 100 5 .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm 8.46 (s, 2H), 8.32, 8.22 (br. s., br.
s., 1H), 7.87-7.50 (m, 2H), 7.30, 6.60 (br. s., br. s., 1H),
7.21-7.16 (m, 1H), 7.09-7.01 (m, 3H), 4.24-4.14 (m, 2H), 4.00-3.94
(m, 9H), 1.25 (d, 3H, J = 6.5 Hz). 102 3 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.44 (s, 1H), 8.30 (s, 2H), 509 8.19 (s, 1H), 7.50
(d, J = 8.5 Hz, 2H), 7.33-7.24 (m, 2H), 7.25-7.15 (m, 3H), 6.87 (d,
J = 8.8 Hz, 2H), 3.79 (br.s, 10H), 2.99 (dd, J = 6.5, 3.5 Hz, 4H),
2.85 (dd, J = 6.3, 3.6 Hz, 4H). 103 3 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.45 (s, 1H), 8.30 (s, 2H), 510 8.19 (s, 1H), 7.52
(d, J = 8.4 Hz, 2H), 7.33-7.12 (m, 5H), 6.90 (d, J = 8.6 Hz, 2H),
3.94-3.66 (m, 14H), 3.03 (t, J = 4.7 Hz, 4H). 105 2 510 101 5
.sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. ppm 8.42 (s, 2H), 8.33,
8.22 (br. s., br. s., 1H), 7.86-7.50 (m, 2H), 7.35, 6.62 (br. s.,
br. s., 1H), 7.21-7.18 (m, 2H), 6.98-6.95 (m, 2H), 4.24-4.13 (m,
2H), 4.00-3.93 (m, 9H), 1.25 (d, 3H, J = 6.0 Hz). 104 3 .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 9.53 (s, 1H), 8.31 (s, 2H), 510 8.24 (s,
1H), 7.44 (s, 1H), 7.32-7.05 (m, 7H), 6.61 (d, J = 7.2 Hz, 1H),
3.86-3.71 (m, 14H), 3.07 (t, J = 4.8 Hz, 4H). 106 2 510 107 2
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.52 (s, 1H), 8.30 (s, 2H),
510 8.21 (s, 1H), 7.57 (d, J = 8.5 Hz, 2H), 7.35-7.12 (m, 5H), 6.87
(d, J = 8.7 Hz, 2H), 4.87 (t, J = 5.6 Hz, 1H), 3.79 (m, 10H),
3.20-3.11 (m, 2H), 3.07-2.89 (m, 3H), 2.12-1.97 (m, 1H), 1.91-1.80
(m, 1H). 108 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.50 (s, 1H),
8.30 (s, 2H), 510 8.20 (s, 1H), 7.55 (d, J = 8.6 Hz, 2H), 7.34-7.12
(m, 5H), 6.84 (d, J = 8.5 Hz, 2H), 4.85-4.68 (m, 1H), 3.79 (d, J =
5.8 Hz, 10H), 3.08-2.95 (m, 1H), 2.94-2.69 (m, 3H), 2.04-1.91 (m,
1H), 1.79-1.67 (m, 1H). 109 1 .sup.1H-NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 9.52 (br. s., 1H), 8.26-8.25 (m, 3H), 7.41-7.36 (m,
3H), 7.24-7.19 (m, 2H), 7.13-7.11 (m, 1H), 6.99 (d, 1H, J = 8.0
Hz), 5.14 (t, 1H, J = 5.5 Hz), 4.54 (d, 2H, J = 5.5 Hz), 3.85-3.78
(m, 12H), 2.93 (t, 2H, J = 5.5 Hz), 2.62 (t, 2H, J = 5.5 Hz). 110 2
.sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. ppm 8.27 (s, 1H), 8.12
(s, 1H), 7.30-7.21 (m, 4H), 7.11 (d, 2H, J = 7.5 Hz), 7.08 (d, 1H,
J = 8.5 Hz), 6.93 (br. s., 1H), 4.50 (s, 2H), 4.02 (s, 2H), 3.92
(br. s., 8H), 3.75 (s, 2H), 3.15 (t, 2H, J = 6.0 Hz), 2.78 (t, 2H,
J = 5.5 Hz). 111 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.33 (s,
1H), 8.31 (s, 2H), 510 8.17 (s, 1H), 7.44 (s, 2H), 7.29 (t, J = 7.5
Hz, 2H), 7.26-7.16 (m, 3H), 6.48 (d, J = 8.3 Hz, 2H), 4.92 (d, J =
3.9 Hz, 1H), 4.45-4.31 (m, 1H), 3.79 (d, J = 10.2 Hz, 10H), 3.39
(dd, J = 10.0, 5.0 Hz, 1H), 3.23 (td, J = 8.6, 3.9 Hz, 1H), 3.04
(d, J = 9.7 Hz, 1H), 2.03 (ddd, J = 13.2, 8.7, 5.0 Hz, 1H), 1.87
(ddt, J = 11.3, 7.0, 3.6 Hz, 1H) - appears to be one peak under
water signal. 112 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.34 (s,
1H), 8.31 (s, 2H), 510
8.17 (s, 1H), 7.44 (s, 2H), 7.33-7.26 (m, 2H), 7.26-7.15 (m, 3H),
6.48 (d, J = 8.5 Hz, 2H), 4.92 (d, J = 3.8 Hz, 1H), 3.79 (d, J =
7.9 Hz, 10H), 3.39 (dd, J = 10.0, 5.0 Hz, 1H), 3.26-3.18 (m, 1H),
3.04 (d, J = 10.0 Hz, 1H), 2.11-1.97 (m, 1H), 1.87 (ddt, J = 11.4,
7.0, 3.7 Hz, 1H) - appears to be one peak under the water signal.
113 1 .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.53 (br,
1H), 8.27, 8.24 (s, s, 3H), 7.40 (s, 1H), 7.36 (d, 1H, J = 8.4 Hz),
7.22-7.16 (m, 2H), 6.97 (t, 2H, J = 7.2 Hz), 6.88 (t, 1H, J = 7.2
Hz), 3.79-3.72 (m, 16H), 2.92 (t, 2H, J = 5.6 Hz), 2.61 (t, 2H, J =
5.6 Hz). 114 5 511 115 1 .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 9.80 (br. s., 1H), 9.50-9.46 (br, 1H), 9.06 (s, 2H),
8.28 (s, 3H), 7.61 (s, 1H), 7.48 (dd, 1H, J = 8.4, 2.0 Hz), 7.16
(d, 1H, J = 8.4 Hz), 7.08-7.00 (m, 2H), 6.78 (d, 1H, J = 7.2 Hz),
6.73-6.69 (m, 1H), 4.27 (br. s., 2H), 3.89-3.37 (m, 10H), 3.39-3.35
(m, 2H), 2.93 (t, 2H, J = 2.0 Hz). 16 3 This spectrum contains some
rotomers in the aromatic region: .sup.1H 512 NMR (400 MHz, DMSO-d6)
.delta. 9.60, 9.49 (s, 1H), 8.30, 8.28 (s, 2H), 8.12, 8.14 (s, 1H),
7.81, 7.77 (s, 1H), 7.48, 7.44 (s, 1H), 7.34-7.10 (m, 5H), 3.95 (d,
J = 7.2 Hz, 2H), 3.79 (t, J = 10.2 Hz, 10H), 2.76 (m, 2H), 2.23 (t,
J = 10.6 Hz, 1H), 2.03-1.76 (m, 2H), 1.56 (br.s, 2H), 1.30 (m, 1H),
1.06 (m, 1H). 117 3 This spectrum contains some rotamers in the
aromatic region: .sup.1H 512 NMR (400 MHz, DMSO-d6) .delta. 9.61,
9.48 (s, 1H), 8.30, 8.26 (s, 2H), 8.14, 8.12 (s, 1H), 7.79, 7.78
(s, 1H), 7.49, 7.44 (s, 1H), 7.34-7.09 (m, 5H), 4.17-3.98 (m, 2H),
3.79 (t, J = 12.4 Hz, 10H), 2.40 (t, J = 6.9 Hz, 2H), 2.24, 2.15
(s, 3H), 1.85 (t, J = 6.7 Hz, 2H). 118 3 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.61, 9.50 (s, s, 1H), 8.32, 8.28 (s, s,
2H), 8.15 (s, 1H), 7.85, 7.82 (s, s, 1H), 7.50, 7.47 (s, s, 1H),
7.29 (t, 2H, J = 7.5 Hz), 7.23 (d, 2H, J = 7.5 Hz), 7.19 (t, 1H, J
= 7.5 Hz), 4.01-3.93 (m, 2H), 3.83-3.77 (m, 12H), 3.24 (t, 2H, J =
11.5 Hz), 2.01 (br. s., 1H), 1.41-1.37 (m, 2H), 1.28-1.20 (m, 2H).
119 4 .sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm 9.62 (br. s., 1H),
513 8.26-8.25 (m, 3H), 7.63 (d, 2H, J = 8.8 Hz), 7.31-7.25 (m, 6H),
7.21-7.17 (m, 1H), 5.12 (d, 1H, J = 4.0 Hz), 4.67-4.64 (m, 1H),
4.49-4.46 (m, 1H), 3.84 (br. s., 4H), 3.80 (br. s., 4H), 3.42-3.39
(m, 2H), 1.64 (s, 6H). 120 4 .sup.1H-NMR (400 MHz, DMSO-d6) .delta.
ppm 9.62 (br. s., 1H), 8.26, 513 8.25 (s, s, 3H), 7.63 (d, 2H, J =
8.4 Hz), 7.33-7.17 (m, 7H), 5.11 (d, 1H, J = 4.4 Hz), 4.68-4.65 (m,
1H), 4.49-4.48 (m, 1H), 3.85-3.80 (m, 8H), 3.43-3.40 (m, 2H), 1.64
(s, 6H). 121 3 .sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm
10.02-9.96 (br, 1H), 513 8.34, 8.33 (s, s, 3H), 7.57 (d, 2H, J =
8.4 Hz), 7.31-7.17 (m, 7H), 4.76-4.41 (m, 4H), 4.28 (s, 1H),
3.86-3.80 (m, 8 H), 1.06 (s, 3H), 0.96 (s, 3H). 122 3 .sup.1H-NMR
(400 MHz, DMSO-d6) .delta. ppm 9.65 (br. s., 1H), 513 8.32 (s, 2H),
8.26 (s, 1H), 7.59 (d, 2H, J = 8.4 Hz), 7.31-7.17 (m, 7H), 5.10 (d,
1H, J = 3.6 Hz), 4.26 (d, 1H, J = 3.6 Hz), 4.21 (s, 1H), 3.85-3.75
(m, 10H), 1.05 (s, 3H), 0.96 (s, 3H). 123 3 This spectrum contains
some rotamers in the aromatic region: .sup.1H 514 NMR (400 MHz,
DMSO-d6) .delta. 9.62, 9.48 (s, 1H), 8.31, 8.27 (s, 2H), 8.14 (s,
1H), 7.81 (s, 1H), 7.48, 7.45 (s, 1H), 7.33-7.12 (m, 4H), 4.21-3.98
(m, 2H), 3.80 (d, J = 12.7 Hz, 10H), 2.84-2.57 (m, 3H), 2.41-2.26
(m, 1H) There is a peak partially obscured by water signal. 124 3
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.63, 9.49 (s, 1H), 8.32,
8.28 (s, 514 2H), 8.15 (s, 1H), 7.82 (s, 1H), 7.49, 7.47 (s, 1H),
7.34-7.26 (m, 2H), 7.26-7.15 (m, 3H), 4.13-4.01 (m, 2H), 3.87-3.64
(m, 13H), 2.74-2.58 (m, 3H), 2.40-2.29 (m, 1H). 125 3 This spectrum
contains some rotamers in the aromatic region: 1H 514 NMR (400 MHz,
DMSO-d6) .delta. 9.64, 9.51 (s, 1H), 8.31, 8.27 (s, 2H), 8.15 (s,
1H), 7.84 (s, 1H), 7.51, 7.47 (s, 1H), 7.35-7.13 (m, 5H), 4.10-3.95
(m, 2H), 3.81 (m, 10H), 3.68-3.55 (m, 2H), 3.17-3.00 (m, 2H), 2.81
(d, J = 12.3 Hz, 1H), 2.70 (t, J = 11.7 Hz, 1H), 2.34 (d, J = 16.8
Hz, 1H). 126 3 This spectrum contains some rotamers in the aromatic
region: 1H 514 NMR (400 MHz, DMSO-d6) .delta. 9.63, 9.51 (s, 1H),
8.31, 8.27 (s, 2H), 8.15 (s, 1H), 7.84 (s, 1H), 7.50, 7.47 (s, 1H),
7.35-7.12 (m, 5H), 4.02 (d, J = 6.2 Hz, 2H), 3.81 (d, J = 13.9 Hz,
10H), 3.65-3.51 (m, 2H), 3.09 (t, J = 9.8 Hz, 1H), 3.00 (d, J = 8.4
Hz, 1H), 2.77 (d, J = 12.3 Hz, 1H), 2.68 (d, J = 11.3 Hz, 1H), 2.34
(d, J = 16.8 Hz, 1H). 127 1 .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 9.79 (br. s., 1H), 9.04 (br. s., 2H), 8.30 (s, 2H),
8.28 (s, 1H), 7.61 (s, 1H), 7.48 (d, 1H, J = 9.6 Hz), 7.33-7.24 (m,
2H), 7.18-7.12 (m, 3H), 4.27 (br. s., 2H), 3.82-3.80 (m, 10H),
3.39-3.35 (m, 2H), 2.93 (t, 2H, J = 1.6 Hz). 128 3 .sup.1H NMR (400
MHz, DMSO-d6) .delta. 9.93 (s, 1H), 8.30 (d, J = 2.9 Hz, 515 2H),
7.90-7.81 (m, 2H), 7.68 (dd, J = 11.1, 8.3 Hz, 2H), 7.33-7.14 (m,
5H), 4.98-4.82 (m, 1H), 4.69-4.38 (m, 4H), 3.78 (s, 2H), 3.15 (s,
2H), 1.62, 1.59 (2 close singlets, 6H), 1.08 (d, J = 6.9 Hz, 3H).
129 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.51 (s, 1H), 8.32 (s,
2H), 515 8.22 (s, 1H), 7.56 (d, J = 8.5 Hz, 2H), 7.37-7.08 (m, 5H),
6.89 (d, J = 8.5 Hz, 2H), 4.92 (d, J = 4.9 Hz, 1H), 4.65 (t, J =
5.6 Hz, 1H), 3.96 (dd, J = 9.5, 4.0 Hz, 1H), 3.80 (h, J = 5.6 Hz,
12H), 3.44 (t, J = 5.5 Hz, 2H). 130 2 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.66 (s, 1H), 8.30 (s, 3H), 515 7.48 (s, 1H),
7.40-7.04 (m, 8H), 3.89-3.88 (m, 0H), 6.58 (d J = 7.9, 1H), 4.96
(s, 1H), 4.67 (s, 1H), 3.97 (dd, J = 9.1, 3.7 Hz, 1H), 3.92-3.67
(m, 14H). 131 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.51 (s,
1H), 8.32 (s, 2H), 515 8.22 (s, 1H), 7.56 (d, J = 8.5 Hz, 2H),
7.37-7.08 (m, 5H), 6.89 (d, J = 8.5 Hz, 2H), 4.92 (d, J = 4.9 Hz,
1H), 4.65 (t, J = 5.6 Hz, 1H), 3.96 (dd, J = 9.5, 4.0 Hz, 1H), 3.80
(h, J = 5.6 Hz, 12H), 3.44 (t, J = 5.5 Hz, 2H). 132 2 .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 9.66 (s, 1H), 8.30 (s, 3H), 515 7.48 (s,
1H), 7.40-7.04 (m, 8H), 3.89-3.88 (m, 0H), 6.58 (d J = 7.9, 1H),
4.96 (s, 1H), 4.67 (s, 1H), 3.97 (dd, J = 9.1, 3.7 Hz, 1H),
3.92-3.67 (m, 14H). 133 3 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 8.21 (s, 3H), 7.76 (s, 1H), 7.57 (s, 1H), 7.32-7.28 (m, 2H),
7.23-7.16 (m, 4H), 4.14-4.11 (m, 2H), 3.88 (br. s., 9H), 3.80 (br.
s., 4H), 3.72-3.69 (m, 2H), 3.62-3.55 (m, 1H), 3.33-3.28 (m, 1H).
134 3 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.21 (s, 3H),
7.75 (s, 1H), 7.59 (s, 1H), 7.35-7.28 (m, 3H), 7.23-7.16 (m, 3H),
4.14-4.11 (m, 2H), 3.93-3.89 (m, 9H), 3.81-3.69 (m, 6H), 3.61-3.55
(m, 1H), 3.33-3.27 (m, 1H). 135 5 .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.64, 9.50 (s, 1H, rotamer), 8.31, 515 8.17
(s, 3H, rotamer), 7.83 (s, 1H), 7.72 (s, 1H), 7.49 (s, 1H), 7.15
(t, J = 7.8 Hz, 2H), 6.79 (d, J = 8.0 Hz, 2H), 6.71 (t, J = 7.5 Hz,
1H), 4.14-4.00 (m, 2H), 3.87-3.64 (m, 10H), 3.47-3.35 (m, 1H),
2.75-2.66 (m, 1H), 2.62 (d, J = 12.4 Hz, 2H), 2.41-2.30 (m, 1H).
136 1 .sup.1H-NMR (400 MHz, DMSO-d.sub.6 + 1d D.sub.2O) .delta. ppm
8.26 (s, 2H), 8.25 (s, 1H), 7.63 (s, 1H), 7.49 (s, 1H), 7.45 (d,
1H, J = 8.4 Hz), 7.41-7.31 (m, 1H), 7.13-7.09 (m, 3H), 4.07 (s,
2H), 3.85-3.72 (m, 10H), 3.18 (d, 2H, J = 6.0 Hz), 2.83 (t, 2H, J =
5.2 Hz). 137 1 .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.58
(br, 1H), 8.32 (s, 2H), 8.25 (s, 1H), 7.43 (s, 1H), 7.38 (d, 1H, J
= 8.0 Hz), 7.33-7.26 (m, 1H), 7.21-7.09 (m, 2H), 7.01 (d, 1H, J =
8.4 Hz), 3.92-3.77 (m, 13H), 2.97 (t, 2H, J = 6.0 Hz), 2.66 (t, 2H,
J = 5.6 Hz). 138 1 .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
9.55 (br, 1H), 8.30 (s, 2H), 8.24 (s, 1H), 7.40-7.35 (m, 3H), 7.21
(t, 1H, J = 10.4 Hz), 7.04 (t, 1H, J = 9.2 Hz), 6.98 (d, 1H, J =
8.0 Hz), 3.80 (br, 12H), 2.92 (t, 2H, J = 5.6 Hz), 2.67-2.61 (m,
2H). 139 1 .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.54
(br, 1H), 8.33 (s, 2H), 8.23 (s, 1H), 7.42-7.29 (m, 4H), 7.10-7.07
(m, 1H), 6.97 (d, 1H, J = 8.4 Hz), 3.80 (br, 13H), 2.91 (t, 2H, J =
5.6 Hz), 2.61 (t, 2H, J = 5.2 Hz). 140 4 .sup.1H-NMR (400 MHz,
DMSO-d6) .delta. ppm 9.65 (br. s., 1H), 8.39 (s, 516 2H), 8.26 (s,
1H), 7.63 (d, 2H, J = 8.4 Hz), 7.46 (dd, 2H, J = 8.4, 5.6 Hz), 7.27
(d, 2H, J = 8.4 Hz), 7.13-7.08 (m, 2H), 5.06 (d, 1H, J = 4.0 Hz),
4.70-4.64 (m, 1H), 3.85-3.70 (m, 8H), 3.45-3.25 (m, 2H), 1.73 (s,
3H), 1.31 (d, 3H, J = 6.4 Hz). 141 1 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.66, 9.52 (s, s, 1H), 8.33 (s, 2H),
8.29, 8.16 (s, s, 1H), 7.83 (s, 1H), 7.50, 7.48 (s, s, 1H),
7.31-7.18 (m, 5H), 4.12-4.03 (m, 2H), 3.84-3.66 (m, 11H), 2.72-2.60
(m, 3H), 2.38-2.33 (m, 1H). 142 1 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.01, 8.77 (s, s, 1H), 8.50 (s, 2H),
8.16, 8.14 (s, s, 1H), 7.55, 7.41 (s, s, 1H), 4.02-3.96 (m, 2H),
3.82-3.64 (m, 10H), 3.34 (br, 2H), 2.73-2.71 (m, 1H), 2.63-2.59 (m,
2H), 2.41-2.36 (m, 1H), 2.17, 2.12 (s, s, 3H). 143 1 .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 10.17 (s, 1H), 8.35 (s, 1H), 519
7.99-7.92 (m, 2H), 7.88-7.81 (m, 2H), 7.63 (s, 1H), 7.28-7.18 (m,
4H), 7.18-7.11 (m, 1H), 3.83 (s, 4H), 3.68 (s, 4H), 3.56 (s, 2H),
3.15 (s, 3H). 145 3 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
9.56 (br. s., 1H), 8.32 (s, 2H), 8.24 (s, 1H), 7.42-7.40 (m, 2H),
7.29 (t, 2H, J = 7.5 Hz), 7.23 (d, 2H, J = 7.0 Hz), 7.20 (t, 1H, J
= 7.5 Hz), 7.01 (d, 1H, J = 8.0 Hz), 3.82-3.80 (m, 10H), 3.67 (s,
2H), 2.82 (t, 2H, J = 5.5 Hz), 2.71 (t, 2H, J = 5.5 Hz), 1.88-1.77
(m, 1H), 0.51-0.47 (m, 2H), 0.41-0.38 (m, 2H). 148 3 .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 9.54, 9.49 (s, 1H), 8.31, 8.27 (s, 522
2H), 8.24 (s, 1H), 7.40 (d, J = 8.2 Hz, 2H), 7.33-7.26 (m, 2H),
7.25-7.16 (m, 3H), 7.01 (d, J = 8.3 Hz, 1H), 3.85-3.76 (m, 10H),
3.59 (s, 2H), 2.90-2.82 (m, 1H), 2.69 (d, J = 16.3 Hz, 4H), 1.05
(d, J = 6.4 Hz, 6H). 149 5 .sup.1H-NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 9.68, 9.55 (s, s, 1H), 8.54 (s, 2H), 8.30, 8.17 (s, s,
1H), 7.96, 7.88 (s, s, 1H), 7.47-7.46 (m, 1H), 7.27-7.25 (m, 2H),
7.19-7.16 (m, 2H), 4.73, 4.67 (s, s, 1H), 3.95-3.88 (m, 10H),
1.06-1.05 (m, 6H). 150 5 .sup.1H-NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 9.70, 9.58 (s, s, 1H), 8.56 (s, 2H), 8.31, 8.18 (s, s,
1H), 7.96, 7.88 (s, s, 1H), 7.48-7.46 (m, 1H), 7.28-7.26 (m, 2H),
7.17-7.14 (m, 1H), 7.04-7.01 (m, 1H), 4.75, 4.68 (s, s, 1H),
4.00-3.89 (m, 10H), 1.06-1.05 (m, 6H). 155 2 .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. ppm 8.28 (s, 1H), 8.20 (s, 2H), 7.53 (d, 2H, J
= 8.4 Hz), 7.48 (d, 2H, J = 8.4 Hz), 7.32-7.26 (m, 2H), 7.23-7.16
(m, 3H), 3.91-3.88 (m, 8H), 3.81 (s, 2H), 1.88-1.71 (m, 8H),
1.65-1.62 (m, 1H), 1.34-1.25 (m, 1H). 156 2 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.44 (s, 1H), 8.30 (s, 2H), 523 8.19 (s, 1H), 7.50
(d, J = 8.4 Hz, 2H), 7.34-7.13 (m, 5H), 6.89 (d, J = 8.6 Hz, 2H),
3.89-3.66 (m, 10H), 3.07 (t, J = 4.9 Hz, 4H), 2.23 (s, 3H) - one
peak is obscured by water signal. 157 2 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.51 (s, 1H), 8.31 (s, 2H), 523 8.24 (s, 1H), 7.43
(s, 1H), 7.31-7.03 (m, 7H), 6.64-6.56 (m, 1H), 3.98-3.65 (m, 12H),
3.11 (t, J = 4.9 Hz, 4H), 2.24 (s, 3H). 158 3 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 8.77 (s, 1H), 8.29 (s, 2H), 523 8.10 (s, 1H), 7.28
(t, J = 7.5 Hz, 2H), 7.25-7.12 (m, 4H), 6.81-6.69 (m, 2H), 3.75 (m,
10H), 3.03 (dd, J = 6.6, 3.5 Hz, 4H), 2.85 (dd, J = 6.2, 3.6 Hz,
4H), 2.12 (s, 3H). 159 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.62 (s, 1H), 8.31 (s, 2H), 524 8.23 (s, 1H), 7.53 (d, J = 8.2 Hz,
2H), 7.35-7.11 (m, 5H), 6.95 (d, J = 8.5 Hz, 2H), 4.87 (br.s, 1H),
4.58 (br.s, 1H), 4.46 (t, J = 15.2 Hz, 4H), 3.78 (s, 2H), 3.77-3.69
(m, 4H), 3.26 (m, 1H), 3.11-3.03 (m, 4H), 1.05 (d, J = 6.4 Hz, 3H).
160 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.66 (s, 1H), 8.31 (d,
J = 1.3 Hz, 524 2H), 8.25 (s, 1H), 7.79 (s, 1H), 7.50 (d, J = 8.1
Hz, 1H), 7.36-7.13 (m, 6H), 6.98-6.87 (m, 1H), 3.89-3.73 (m, 10H),
3.64-3.52 (m, 4H), 3.42 (s, 2H), 2.42-2.28 (m, 9H). 161 2 .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 9.51 (s, 1H), 8.30 (s, 2H), 524 8.21
(s, 1H), 7.55 (d, J = 8.5 Hz, 2H), 7.33-7.24 (m, 2H), 7.24-7.15 (m,
3H), 6.90 (d, J = 9.0 Hz, 2H), 4.25-4.18 (m, 1H), 3.78
(br.s, J = 6.9 Hz, 10H), 3.15-3.05 (m, 1H), 2.86-2.74 (m, 1H),
2.62-2.54 (m, 1H), 2.05-1.91 (m, 2H), 1.72-1.65 (m, 1H), 1.56-1.36
(m, 2H). This spectra has slight evidence of rotamers in the
aromatic region. 162 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.51
(s, 1H), 8.30 (s, 2H), 524 8.21 (s, 1H), 7.55 (d, J = 8.5 Hz, 2H),
7.33-7.24 (m, 2H), 7.24-7.15 (m, 3H), 6.90 (d, J = 9.0 Hz, 2H),
4.25-4.18 (m, 1H), 3.78 (br.s, J = 6.9 Hz, 10H), 3.15-3.05 (m, 1H),
2.86-2.74 (m, 1H), 2.62-2.54 (m, 1H), 2.05-1.91 (m, 2H), 1.72-1.65
(m, 1H), 1.56-1.36 (m, 2H). This spectra has slight evidence of
rotamers in the aromatic region. 163 3 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.90 (s, 1H), 9.74 (s, 1H), 524 8.31 (s, 2H), 8.27
(s, 1H), 7.58 (s, 1H), 7.56-7.50 (m, 1H), 7.32-7.25 (m, 2H),
7.25-7.11 (m, 4H), 5.42-5.28 (m, 1H), 4.59-4.25 (m, 4H), 3.92-3.71
(m, 12H), 3.09-2.94 (m, 2H). 164 3 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.62 (s, 1H), 8.39-8.06 (m, 524 3H), 7.62-6.91 (m, 7H),
4.93 (d, J = 23.8 Hz, 2H), 4.58 (dd, J = 29.9, 13.0 Hz, 4H), 4.02
(s, 2H), 3.78 (s, 2H), 3.57-2.85 (m, 3H), 2.76 (t, J = 5.8 Hz, 2H),
1.64 (bs, 2H). 165 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.43
(s, 1H), 8.30 (s, 2H), 524 8.19 (s, 1H), 7.49 (d, J = 8.2 Hz, 2H),
6.87 (d, J = 8.4 Hz, 2H), 4.78 (s, 1H), 3.79 (p, J = 7.1 Hz, 10H),
3.62-3.54 (m, 1H), 3.51 (dd, J = 11.7, 4.0 Hz, 1H), 2.64-2.54 (m,
2H), 2.48-2.37 (m, 1H), 1.86 (dt, J = 12.1, 4.1 Hz, 1H), 1.73 (dt,
J = 13.5, 3.8 Hz, 1H), 1.60-1.44 (m, 2H), 1.30-1.16 (m, 2H); one
signal is partially obscured by water peak. 166 2 .sup.1H NMR (400
MHz, DMSO-d6) .delta. 9.43 (s, 1H), 8.30 (s, 2H), 524 8.19 (s, 1H),
7.49 (d, J = 8.4 Hz, 2H), 6.87 (d, J = 8.4 Hz, 2H), 4.77 (s, 1H),
3.65-3.43 (m, 2H), 2.57 (t, J = 10.7 Hz, 1H), 2.42 (t, J = 10.2 Hz,
1H), 1.87 (dq, J = 12.6, 4.3 Hz, 1H), 1.73 (dt, J = 13.2, 3.8 Hz,
1H), 1.62-1.44 (m, 1H), 1.31-1.15 (m, 2H). 167 2 .sup.1H NMR (400
MHz, DMSO-d6) .delta. 9.32 (s, 1H), 8.30 (s, 2H), 524 8.15 (s, 1H),
7.43 (s, 2H), 7.34-7.12 (m, 5H), 6.55 (d, J = 8.5 Hz, 2H), 4.70
(dd, J = 6.3, 5.1 Hz, 1H), 3.78 (m, 10H), 3.60 (dt, J = 8.4, 4.5
Hz, 1H), 3.48 (dt, J = 9.3, 4.4 Hz, 1H), 3.15 (ddd, J = 10.7, 8.6,
6.5 Hz, 1H), 3.04-2.92 (m, 1H), 2.04-1.77 (m, 4H) - one peak is
obscured by water signal. 168 2 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.32 (s, 1H), 8.30 (s, 2H), 524 8.15 (s, 1H), 7.43 (s, 2H),
7.35-7.13 (m, 5H), 6.55 (d, J = 8.5 Hz, 2H), 4.70 (dd, J = 6.4, 5.1
Hz, 1H), 3.78 (m, 10H), 3.66-3.56 (m, 2H), 3.48 (dt, J = 9.5, 4.4
Hz, 1H), 3.20-3.09 (m, 1H), 3.04-2.91 (m, 2H), 2.04-1.81 (m, 4H)
one peak is obscured by water signal. 169 2 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.50 (s, 1H), 8.30 (s, 2H), 524 8.21 (s, 1H),
7.61-7.50 (m, 2H), 7.34-7.12 (m, 5H), 6.87 (d, J = 8.7 Hz, 2H),
3.87-3.69 (m, 10H), 2.86-2.72 (m, 2H), 1.88-1.76 (m, 1H), 1.74-1.55
(m, 2H), 1.49-1.35 (m, 1H) - one peak obscured by water signal. 170
2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.50 (s, 1H), 8.30 (s,
2H), 524 8.21 (s, 1H), 7.55 (d, J = 8.0 Hz, 2H), 7.34-7.12 (m, 4H),
6.87 (d, J = 8.7 Hz, 2H), 3.87-3.69 (m, 10H), 2.86-2.73 (m, 2H),
1.89-1.75 (m, 1H), 1.74-1.55 (m, 2H), 1.49-1.35 (m, 1H) - one peak
obscured by water signal. 171 2 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.60 (s, 1H), 8.35 (s, 1H), 525 8.32 (s, 2H), 8.24 (s, 1H),
7.94 (dd, J = 8.9, 2.7 Hz, 1H), 7.36-7.15 (m, 5H), 6.75 (d, J = 8.9
Hz, 1H), 4.84 (dt, J = 8.9, 4.7 Hz, 1H), 3.79 (d, J = 5.4 Hz, 10H),
3.11 (dd, J = 12.0, 3.9 Hz, 1H), 2.77 (dt, J = 12.2, 4.0 Hz, 1H),
2.04 (d, J = 11.0 Hz, 1H), 1.66 (dt, J = 12.5, 4.3 Hz, 1H),
1.57-1.37 (m, 2H) - one peak obscured by DMSO signal. 172 3 .sup.1H
NMR (400 MHz, DMSO-d6) .delta. ppm 9.67 (s, 1H), 8.32 (s, 2H), 525
8.26 (s, 1H), 7.65 (s, 1H), 7.63 (d, 1H, J = 8.4 Hz), 7.33-7.15 (m,
7H), 5.48 (d, 1H, J = 3.6 Hz), 4.77-4.65 (m, 3H), 4.10 (d, 1H, J =
5.2 Hz), 4.01 (d, 1H, J = 5.2 Hz), 3.88-3.70 (m, 10H), 1.11 (s,
3H). 173 4 .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.67,
9.54 (s, s, 1H), 8.34 (s, 2H), 8.30, 8.17 (s, s, 1H), 7.83 (s, 1H),
7.50, 7.47 (s, s, 1H), 7.41-7.34 (m, 5H), 5.51 (s, 1H), 5.43 (s,
1H), 4.12-4.03 (m, 2H), 3.91-3.87 (m, 8H), 3.72-3.66 (m, 2H),
3.41-3.37 (m, 1H), 2.72-2.59 (m, 3H), 2.37-2.33 (m, 2H). 174 3
rotamers in aromatic region: 1H NMR (400 MHz, DMSO-d6) .delta. 526
9.60 (s, 1H), 8.31 (s, 2H), 8.14 (s, 1H), 7.82, 7.80 (s, 1H), 7.47,
7.45 (s, 1H), 7.33-7.13 (m, 5H), 4.04-3.89 (m, 2H), 3.80 (m, 8H),
2.76 (m, 2H), 2.16 (s, 3H), 1.85 (m, 2H), 1.75 (m, 1H), 1.43 (m,
2H), 1.21 (m, 2H). 175 3 527 176 3 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.66, 9.53 (s, s, 1H), 8.32 (s, 2H),
8.29, 8.15 (s, s, 1H), 7.87, 7.85 (s, s, 1H), 7.46 (s, 1H),
7.31-7.17 (m, 5H), 4.31-4.12 (m, 2H), 3.99-3.79 (m, 11H), 2.69-2.55
(m, 3H), 2.29-2.22 (m, 5H), 2.10-2.05 (m, 1H). 177 3 .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 9.67, 9.54 (s, s, 1H),
8.31-8.28 (m, 2H), 8.15 (s, 1H), 7.88 (s, 1H), 7.47 (s, 1H),
7.30-7.17 (m, 5H), 4.32-4.22 (m, 2H), 3.83-3.79 (m, 11H), 2.84-2.64
(m, 4H), 2.44-2.28 (m, 5H), 2.18-2.13 (m, 1H). 178 4 .sup.1H-NMR
(500 MHz, DMSO-d.sub.6) .delta. ppm 9.70, 9.57 (s, s, 1H), 8.77 (s,
2H), 8.32, 8.19 (s, s, 1H), 7.84 (s, 1H), 7.75 (d, 2H, J = 7.5 Hz),
7.69 (t, 1H, J = 7.5 Hz), 7.58 (t, 2H, J = 8.0 Hz), 7.51, 7.48 (s,
s, 1H), 4.16-3.95 (m, 6H), 3.93-3.87 (m, 4H), 3.73-3.64 (m, 2H),
3.39-3.31 (m, 1H), 2.72-2.55 (m, 3H), 2.38-2.34 (m, 2H). 179 1 528
180 3 This spectrum contains some rotomers in the aromatic region:
.sup.1H 528 NMR (400 MHz, DMSO-d6) .delta. 9.63, 9.51 (s, 1H),
8.31, 8.27 (s, 2H), 8.15 (s, 1H), 7.83 (s, 1H), 7.49 (s, 1H),
7.34-7.14 (m, 5H), 4.20-4.08 (m, 2H), 3.79 (s, 10H), 3.58-3.41 (m,
2H), 2.29-2.09 (m, 3H) - some peaks are partially obscured by water
signal. 181 3 This spectrum contains some rotomers in the aromatic
region: .sup.1H 528 NMR (300 MHz, DMSO-d6) .delta. 9.65, 9.51 (s,
1H), 8.32, 8.28 (s, 2H), 8.16 (s, 1H), 7.84 (s, 1H), 7.50 (s, 1H),
7.35-7.13 (m, 5H), 4.22-4.04 (m, 2H), 3.81 (d, J = 9.4 Hz, 10H),
3.46 (m, 1H), 2.58 (t, J = 11.8 Hz, 2H), 2.15 (s, 3H), 2.03-1.87
(m, 2H), 1.72 (t, J = 10.6 Hz, 2H). 182 3 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.65, 9.63 (s, 1H), 8.30, 8.31 (s, 528 2H), 8.14
(d, J = 2.9 Hz, 1H), 7.91 (s, 1H), 7.46, 7.43 (s, 1H), 7.34-7.10
(m, 5H), 4.09-3.93 (m, 2H), 3.90-3.67 (m, 10H), 2.90-2.69 (m, 2H),
2.33-2.23 (m, 2H), 1.52-1.24 (m, 4H). 183 3 This spectrum contains
some rotomers in the aromatic region: .sup.1H 528 NMR (400 MHz,
DMSO-d6) .delta. 9.62, 9.46 (s, 1H), 8.30, 8.26 (s, 2H), 8.14 (s,
1H), 7.82 (d, J = 2.7 Hz, 1H), 7.46 (s, 1H), 7.35-7.12 (m, 4H),
5.01-4.38 (m, 5H), 4.16-3.96 (m, 2H), 3.78 (s, 2H), 3.75-3.60 (m,
2H), 3.38 (dd, J = 11.6, 8.3 Hz, 1H), 3.21-2.99 (m, 2H), 2.76-2.56
(m, 3H), 2.40-2.27 (m, 1H), 1.05 (d, J = 6.4 Hz, 3H). 184 3
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.06, 8.79 (s, s,
1H), 8.32 (s, 2H), 8.18, 8.16 (s, s, 1H), 7.79, 7.70 (s, s, 1H),
7.29 (t, 2H, J = 9.0 Hz), 7.24-7.17 (m, 3H), 4.03-3.94 (m, 2H),
3.79-3.63 (m, 7H), 3.43-3.38 (m, 1H), 2.83-2.56 (m, 3H), 2.38-2.32
(m, 1H), 2.12, 2.07 (s, s, 3H). 185 3 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 8.97, 8.73 (s, s, 2H), 8.30, 8.14 (s, s,
3H), 7.54, 7.40 (s, s, 1H), 7.31-7.17 (m, 4H), 4.02-3.98 (m, 2H),
3.79-3.64 (m, 10H), 3.78-3.35 (m, 1H), 2.74-2.59 (m, 4H), 2.49-2.36
(m, 2H), 2.16-2.12 (m, 3H). 186 1 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.62, 9.50 (s, s, 1H), 8.32-8.14 (m, 3H),
7.83-7.81 (m, 1H), 7.48-7.46 (m, 1H), 7.29-7.16 (m, 5H), 4.10-4.02
(m, 3H), 3.82-3.64 (m, 11H), 2.71-2.58 (m, 3H), 2.49-2.33 (m, 1H),
1.56 (d, 3H, J = 7.2 Hz). 187 5 511 188 3 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.67 (s, 1H), 8.31 (s, 2H), 530 8.26 (s, 1H), 7.76
(s, 1H), 7.54 (d, J = 8.0 Hz, 1H), 7.35-7.12 (m, 6H), 6.92 (d, J =
7.5 Hz, 1H), 3.94-3.51 (m, 16H). 189 4 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.66, 9.52 (s, s, 1H), 8.33 (s, 2H),
8.28, 8.16 (s, s, 1H), 7.83 (s, 1H), 7.49, 7.47 (s, s, 1H), 7.39
(d, 2H, J = 7.5 Hz), 7.34 (t, 2H, J = 7.5 Hz), 7.24 (t, 1H, J = 7.0
Hz), 5.96 (d, 1H, J = 4.5 Hz), 5.66 (d, 1H, J = 4.0 Hz), 4.14-4.02
(m, 2H), 3.85-3.75 (m, 8H), 3.73-3.62 (m, 2H), 3.41-3.39 (m, 1H),
2.71-2.55 (m, 3H), 2.35-2.31 (m, 2H). 190 2 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.64, 9.51 (s, 1H, rotamer), 530 8.51 (s, 2H),
7.83 (s, 1H), 8.17 (s, 1H) 7.49 (s, 1H), 7.35-7.26 (m, 2H),
7.05-6.91 (m, 3H), 4.96 (s, 2H), 4.10-4.03 (m, 2H), 3.86 (m, 8H),
3.76-3.64 (m, 2H), 3.45-3.27 (m, 1H), 2.70-2.62 (m, 3H), 2.41-2.30
(m, 2H). 191 3 This spectrum contains some rotomers in the aromatic
region: .sup.1H 530 NMR (400 MHz, DMSO-d6) .delta. 9.65, 9.53 (s,
1H), 8.31, 8.28 (s, 2H), 8.15 (s, 1H), 7.84 (s, 1H), 7.50, 7.48 (s,
1H), 7.32-7.13 (m, 5H), 4.32 (d, J = 21.6 Hz, 2H), 3.79 (m, 10H),
2.84-2.60 (m, 4H), 1.67-1.43 (m, 4H). 192 3 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.75 (br. s., 1H), 8.33 (s, 2H), 8.29 (s,
1H), 7.63 (s, 1H), 7.54 (dd, 1H, J = 8.0, 1.5 Hz), 7.37 (br. s.,
1H), 7.30 (t, 2H, J = 7.5 Hz), 7.24 (d, 2H, J = 7.5 Hz), 7.20 (t,
1H, J = 7.0 Hz), 7.08 (d, 1H, J = 8.5 Hz), 4.42 (s., 2H), 4.31 (s,
2H), 3.88-3.75 (m, 10H). 193 4 .sup.1H-NMR (400 MHz, DMSO-d6)
.delta. ppm 9.63 (br. s., 1H), 8.39 (s, 530 2H), 8.25 (s, 1H), 7.61
(d, 2H, J = 8.4 Hz), 7.46 (dd, 2H, J = 8.4, 5.6 Hz), 7.39 (d, 2H, J
= 8.8 Hz), 7.13-7.08 (m, 2H), 4.93 (s, 1H), 3.85-3.70 (m, 8H),
2.67-2.64 (m, 2H), 1.73 (s, 3H), 1.41 (s, 6H). 194 1 .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 9.64, 9.49 (s, br. s., 1H),
8.28, 8.14 (s, s, 1H), 7.96 (s, 1H), 7.83 (br. s., 1H), 7.48 (br.
s., 1H), 6.80-6.75 (br, 1H), 4.91-4.80 (m, 1H), 4.76-4.45 (m, 3H),
4.09-3.99 (m, 2H), 3.75-3.63 (m, 2H), 3.26-2.99 (m, 4H), 2.72-2.62
(m, 3H), 2.42-2.33 (m, 2H), 1.04 (br. s., 3H). 195 4 .sup.1H-NMR
(400 MHz, DMSO-d6) .delta. ppm 9.67 (s, 1H), 8.39 (s, 2H), 532 8.26
(s, 1H), 7.63 (d, 2H, J = 8.0 Hz), 7.48-7.44 (m, 2H), 7.26 (d, 2H,
J = 8.0 Hz), 7.13-7.09 (m, 2H), 5.16 (d, 1H, J = 4.0 Hz), 4.71 (t,
1H, J = 5.6 Hz), 4.50-4.46 (m, 1H), 3.83-3.82 (m, 8H), 3.42-7.39
(m, 2H), 2.50 (s, 3H), 1.72 (s, 3H). 196 1 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.63, 9.50 (s, s, 1H), 8.32-8.16 (m, 3H),
7.82 (s, 1H), 7.49-7.47 (m, 1H), 7.29-7.26 (m, 2H), 7.14-7.09 (m,
2H), 4.11-4.02 (m, 2H), 3.83-3.65 (m, 12H), 3.42-3.39 (m, 1H),
2.71-2.59 (m, 3H), 2.38-2.32 (m, 1H). 197 1 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.64, 9.50 (s, s, 1H), 8.34-8.16 (m, 3H),
7.83 (s, 1H), 7.50-7.47 (m, 1H), 7.36-7.30 (m, 1H), 7.11-7.00 (m,
3H), 4.10-4.03 (m, 2H), 3.83-3.65 (m, 12H), 3.39-3.33 (m, 1H),
2.71-2.60 (m, 3H), 2.38-2.33 (m, 1H). 198 1 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.64, 9.50 (s, s, 1H), 8.30 (s, 2H), 8.16
(s, 1H), 7.83 (s, 1H), 7.49-7.47 (m, 1H), 7.34-7.26 (m, 2H),
7.19-7.13 (m, 2H), 4.11-4.03 (m, 2H), 3.83-3.66 (m, 12H), 3.42-3.34
(m, 1H), 2.71-2.50 (m, 3H), 2.38-2.32 (m, 1H). 199 5 .sup.1H-NMR
(500 MHz, DMSO-d.sub.6) .delta. ppm 9.67, 9.54 (s, s, 1H), 8.54 (s,
2H), 8.30, 8.17 (s, s, 1H), 7.83 (s, 1H), 7.50, 7.47 (s, s, 1H),
7.33-7.30 (m, 2H), 7.21-7.16 (m, 3H), 4.14-4.03 (m, 2H), 3.94-3.88
(m, 8H), 3.72-3.64 (m, 2H), 3.41-3.36 (m, 1H), 2.71-2.58 (m, 3H),
2.37-2.31 (m, 1H). 200 5 1H NMR (400 MHz, DMSO-d6) .delta. 9.50,
9.64 (s, 1H, rotamer), 533 8.29, 8.17 (s, 2H, rotamer), 7.83 (s,
1H), 7.68 (s, 1H), 7.48, 7.51 (s, 1H, rotamer), 7.00 (t, J = 8.8
Hz, 2H), 6.80 (dd, J = 8.9, 4.6 Hz, 2H), 4.23-3.50 (m, 12H),
3.49-3.37 (m, 2H), 2.82-2.55 (m, 3H), 2.42-2.17 (m, 2H). 201 4
.sup.1H-NMR (400 MHz, CD3OD) .delta. ppm 8.23 (s, 2H), 8.13 (s,
1H), 533 8.05 (s, 1H), 7.56 (s, 1H), 7.32-7.29 (m, 2H), 7.06-7.01
(m, 2H),
4.09-4.06 (m, 2H), 3.93-3.87 (m, 8H), 1.68 (3, 3H), 1.19 (s, 3H).
202 5 .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.64, 9.50
(s, s, 1H), 8.34 (s, 2H), 8.29, 8.16 (s, s, 1H), 7.82 (s, 1H),
7.50, 7.49 (s, s, 1H), 7.20-7.16 (m, 2H), 7.05-7.01 (m, 2H),
4.10-4.00 (m, 2H), 3.86-3.78 (m, 8H), 3.75-3.61 (m, 2H), 3.47-3.36
(m, 1H), 2.71-2.62 (m, 3H), 2.40-2.30 (m, 1H). 203 5 .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 9.65, 9.52 (s, s, 1H), 8.37 (s,
2H), 8.28, 8.16 (s, s, 1H), 7.82 (s, 1H), 7.50, 7.46 (s, s, 1H),
7.39-7.33 (m, 1H), 7.16-7.11 (m, 2H), 7.06-7.02 (m, 1H), 4.18-4.01
(m, 2H), 3.86-3.80 (m, 8H), 3.71-3.65 (m, 2H), 3.39-3.37 (m, 2H),
2.71-2.56 (m, 3H), 2.34-2.30 (m, 1H). 204 5 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.64, 9.52 (br. s., br. s., 1H), 8.38 (s,
2H), 8.32, 8.16 (s, s, 1H), 7.82 (s, 1H), 7.50, 7.48 (s, 1H),
7.39-7.35 (m, 1H), 6.95-6.85 (m, 2H), 6.81 (dd, 1H, J = 8.0, 2.0
Hz), 4.10-4.06 (m, 2H), 3.87-3.82 (m, 8H), 3.76-3.61 (m, 2H),
3.45-3.35 (m, 1H), 2.66-2.62 (m, 3H), 2.41-2.29 (m, 1H). 205 5
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 10.14 (br. s., 1H),
8.56 (s, 2H), 8.36 (s, 1H), 7.77-7.75 (m, 2H), 7.42-7.40 (m, 2H),
7.28-7.25 (m, 2H), 7.18-7.14 (m, 1H), 7.07-7.03 (m, 1H), 3.91 (br.
s., 10H), 3.25-3.23 (m, 1H), 3.12-3.06 (m, 1H), 1.13 (d, 3H, J =
6.0 Hz). 206 5 .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
10.08 (br. s., 1H), 8.56 (s, 2H), 8.35 (s, 1H), 7.77-7.75 (m, 2H),
7.36-7.25 (m, 4H), 7.18-7.14 (m, 1H), 7.06-7.03 (m, 1H), 3.91 (br.
s., 10H), 3.25-3.22 (m, 1H), 3.11-3.06 (m, 1H), 1.13 (d, 3H, J =
6.0 Hz). 207 4 .sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm 9.66,
9.53 (s, s, 1H), 535 8.39 (s, 2H), 8.30, 8.16 (s, s, 1H), 7.94,
7.87 (s, s, 1H), 7.48-7.44 (m, 3H), 7.15-7.10 (m, 2H), 5.89 (s,
1H), 4.73, 4.67 (s, s, 1H), 4.00, 3.94 (s, s, 2H), 3.83-3.79 (m,
8H), 1.81 (s, 3H), 1.06 (s, 6H). 208 4 .sup.1H-NMR (400 MHz,
DMSO-d6) .delta. ppm 9.66, 9.52 (s, s, 1H), 535 8.39 (s, 2H), 8.28,
8.15 (s, s, 1H), 7.93, 7.86 (s, s, 1H), 7.47-7.43 (m, 3H),
7.15-7.01 (m, 2H), 5.89 (s, 1H), 4.73, 4.67 (s, s, 1H), 4.00, 3.94
(s, s, 2H), 3.82-3.78 (m, 8H), 1.81 (s, 3H), 1.06 (s, 6H). 211 3
537 212 3 .sup.1NMR (400 MHz, DMSO-d6) .delta. 8.77 (s, 1H), 8.29
(s, 2H), 537 8.09 (s, 1H), 7.32-7.24 (m, 2H), 7.24-7.12 (m, 4H),
6.80-6.68 (m, 2H), 3.75 (m, 10H), 3.13-3.03 (m, 4H), 2.43 (t, J =
5.0 Hz, 4H), 2.21 (s, 3H), 2.12 (s, 3H). 215 3 .sup.1H NMR (400
MHz, DMSO-d6) .delta. 9.55 (s, 1H), 8.30 (s, 2H), 538 8.23 (s, 1H),
7.39 (d, J = 9.4 Hz, 1H), 7.33-7.13 (m, 6H), 7.00 (d, J = 8.1 Hz,
1H), 3.80 (d, J = 8.1 Hz, 10H), 3.60-3.47 (m, 4H), 3.29 (s, 4H),
3.26 (s, 3H), 2.76-2.59 (m, 4H). 216 2 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.51 (s, 1H), 8.30 (s, 2H), 538 8.21 (s, 1H), 7.55
(d, J = 8.6 Hz, 2H), 7.34-7.13 (m, 5H), 6.90 (d, J = 8.5 Hz, 2H),
4.34-4.27 (m, 1H), 3.79 (m, 10H), 2.91-2.83 (m, 1H), 2.61-2.52 (m,
1H), 2.20 (s, 3H), 2.08-1.87 (m, 3H), 1.75-1.67 (m, 1H), 1.58-1.49
(m, 1H), 1.37-1.28 (m, 1H). 217 2 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 9.51 (s, 1H), 8.30 (s, 2H), 538 8.21 (s, 1H), 7.55 (d, J =
8.6 Hz, 2H), 7.34-7.13 (m, 5H), 6.90 (d, J = 8.5 Hz, 2H), 4.34-4.27
(m, 1H), 3.79 (m, 10H), 2.91-2.83 (m, 1H), 2.61-2.52 (m, 1H), 2.20
(s, 3H), 2.08-1.87 (m, 3H), 1.75-1.67 (m, 1H), 1.58-1.49 (m, 1H),
1.37-1.28 (m, 1H). 218 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.50 (s, 1H), 8.30 (s, 2H), 538 8.21 (s, 1H), 7.55 (d, J = 8.7 Hz,
2H), 7.34-7.14 (m, 5H), 6.89 (d, J = 8.5 Hz, 2H), 3.94 (dd, J =
9.6, 5.4 Hz, 1H), 3.79 (d, J = 3.9 Hz, 10H), 2.97 (br.s, 1H), 2.37
(s, 3H), 2.21 (br.s, 1H), 2.03-1.88 (m, 1H), 1.76-1.63 (m, 2H),
1.64-1.51 (m, 1H) - a peak is obscured by the water signal. 219 2
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.50 (s, 1H), 8.30 (s, 2H),
538 8.21 (s, 1H), 7.56 (d, J = 8.6 Hz, 2H), 7.35-7.12 (m, 5H), 6.89
(d, J = 8.5 Hz, 2H), 3.94 (dd, J = 9.6, 5.4 Hz, 1H), 3.79 (d, J =
6.6 Hz, 10H), 2.98 (br.s, 1H), 2.38 (br.s, 3H), 2.22 (s, 1H),
2.03-1.89 (m, 1H), 1.77-1.64 (m, 2H), 1.64-1.53 (m, 1H) a peak is
obscured by the water signal. 220 3 .sup.1H-NMR (500 MHz,
CD.sub.3OD) .delta. ppm 8.25 (s, 2H), 8.19 (s, 1H), 7.38 (d, 1H, J
= 8.0 Hz), 7.37 (s, 1H), 7.32 (d, 1H, J = 8.0 Hz), 7.31 (d, 1H, J =
8.0 Hz), 7.24-7.20 (m, 3H), 7.10 (d, 1H, J = 8.0 Hz), 4.64 (s, 2H),
4.11-4.06 (m, 1H), 3.93-3.90 (m, 4H), 3.87-3.84 (m, 6H), 3.78-3.76
(m, 2H), 2.93-2.87 (m, 4H), 2.62-2.54 (m, 2H), 1.22 (d, 3H, J = 6.0
Hz). 223 1 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.42 (s, 1H),
8.31 (s, 2H), 538 8.19 (s, 1H), 7.49 (d, J = 8.4 Hz, 2H), 7.35-7.22
(m, 4H), 7.17 (tt, J = 5.5, 2.5 Hz, 1H), 6.86 (d, J = 8.6 Hz, 2H),
4.77 (d, J = 4.7 Hz, 1H), 4.05 (q, J = 7.3 Hz, 1H), 3.78 (q, J =
7.0, 6.4 Hz, 8H), 3.62-3.47 (m, 2H), 3.43-3.35 (m, 1H), 2.62-2.52
(m, 1H), 2.42 (dd, J = 11.4, 9.0 Hz, 1H), 1.86 (dt, J = 12.5, 4.1
Hz, 1H), 1.73 (dt, J = 13.0, 3.8 Hz, 1H), 1.56 (d, J = 7.3 Hz, 4H),
1.29-1.17 (m, 1H). 224 1 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.43 (s, 1H), 8.31 (s, 2H), 538 8.19 (s, 1H), 7.49 (d, J = 8.3 Hz,
2H), 7.36-7.23 (m, 4H), 7.23-7.13 (m, 1H), 6.86 (d, J = 8.6 Hz,
2H), 4.77 (d, J = 4.7 Hz, 1H), 4.05 (q, J = 7.2 Hz, 1H), 3.78 (m,
8H), 3.64-3.46 (m, 2H), 2.56 (td, J = 11.7, 3.0 Hz, 1H), 2.42 (dd,
J = 11.4, 9.0 Hz, 1H), 1.87 (m, 1H), 1.73 (m, 1H), 1.63-1.45 (m,
4H), 1.22 (tdd, J = 12.1, 9.5, 4.1 Hz, 1H). 225 2 .sup.1H NMR (400
MHz, DMSO-d6) .delta. 9.42 (s, 1H), 8.30 (s, 2H), 538 8.18 (s, 1H),
7.48 (d, J = 8.6 Hz, 2H), 7.24 (tt, J = 19.9, 7.5 Hz, 5H), 6.85 (d,
J = 9.3 Hz, 2H), 4.38 (s, 1H), 3.98-3.65 (m, 10H), 2.97 (d, J = 6.1
Hz, 2H), 2.89-2.80 (m, 2H), 1.78 (m, 1H), 1.57-1.40 (m, 3H), 1.16
(s, 3H). 226 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.42 (s, 1H),
8.31 (s, 2H), 539 8.19 (s, 1H), 7.48 (d, J = 8.4 Hz, 2H), 7.34-7.14
(m, 5H), 6.86 (d, J = 8.5 Hz, 2H), 4.38 (s, 1H), 3.79 (s, 10H),
2.98 (d, J = 6.5 Hz, 2H), 2.90-2.79 (m, 2H), 1.79 (dt, J = 11.4,
6.1 Hz, 2H), 1.59-1.52 (m, 1H), 1.47 (t, J = 6.1 Hz, 2H), 1.16 (s,
3H). 227 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.42 (s, 1H),
8.31 (s, 2H), 538 8.19 (s, 1H), 7.48 (d, J = 8.4 Hz, 2H), 7.33-7.15
(m, 5H), 6.86 (d, J = 8.3 Hz, 2H), 4.38 (s, 1H), 3.79 (br.s, 10H),
2.97 (dt, J = 7.4, 3.9 Hz, 2H), 2.89-2.80 (m, 2H), 1.79 (dt, J =
11.4, 5.9 Hz, 1H), 1.54 (dt, J = 13.5, 6.6 Hz, 1H), 1.47 (t, J =
6.0 Hz, 2H), 1.16 (s, 3H). 228 1 .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 8.74 (s, 1H), 8.09 (s, 1H), 538 7.63 (s, 1H), 7.30-7.11 (m,
6H), 6.81-6.67 (m, 2H), 6.29 (s, 2H), 3.68 (m, 10H), 3.01 (dd, J =
6.6, 3.5 Hz, 4H), 2.84 (dd, J = 6.3, 3.6 Hz, 4H), 2.12 (s, 3H). 229
1 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.53 (s, 1H), 8.22 (s,
1H), 539 7.64 (s, 1H), 7.40 (d, J = 7.2 Hz, 2H), 7.31-7.13 (m, 5H),
7.01 (d, J = 8.4 Hz, 1H), 6.31 (s, 2H), 4.47 (br.s, 1H), 3.83-3.53
(m, 14H), 2.73 (br.s, 4H), 2.57 (br.s, 2H). 221 3 .sup.1H-NMR (500
MHz, DMSO-d.sub.6) .delta. ppm 9.58 (br. s., 1H), 8.32 (s, 2H),
8.24 (s, 1H), 7.41 (d, 1H, J = 7.5 Hz), 7.40 (s, 1H), 7.29 (d, 2H,
J = 7.5 Hz), 7.23 (d, 2H, J = 7.5 Hz), 7.19 (d, 1H, J = 7.5 Hz),
7.01 (d, 1H, J = 7.5 Hz), 4.39 (d, 1H, J = 4.0 Hz), 3.89-3.79 (m,
11H), 3.56 (s, 2H), 2.74-2.67 (m, 4H), 2.44-2.31 (m, 2H), 1.07 (d,
3H, J = 5.5 Hz). 222 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.44
(s, 1H), 8.31 (s, 2H), 538 8.19 (s, 1H), 7.51 (s, 2H), 7.34-7.27
(m, 2H), 7.27-7.16 (m, 3H), 6.88 (d, J = 8.5 Hz, 2H), 4.95-4.73 (m,
2H), 4.47 (dd, J = 24.2, 13.2 Hz, 2H), 4.34 (d, J = 4.2 Hz, 2H),
3.78 (d, J = 10.8 Hz, 3H), 3.66-3.47 (m, 2H), 3.40 (d, J = 12.1 Hz,
1H), 3.27-3.09 (m, 2H), 2.59 (d, J = 10.0 Hz, 1H), 2.43 (t, J =
10.2 Hz, 1H), 1.88 (d, J = 12.4 Hz, 1H), 1.73 (dd, J = 11.1, 7.2
Hz, 1H), 1.53 (d, J = 12.8 Hz, 1H), 1.31-1.17 (m, 2H), 1.04 (d, J =
6.3 Hz, 10H). 231 1 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.41 (s,
1H), 8.19 (s, 1H), 539 7.65 (s, 1H), 7.50 (d, J = 8.4 Hz, 2H),
7.35-7.11 (m, 4H), 6.87 (d, J = 8.5 Hz, 2H), 6.31 (s, 2H), 4.78 (d,
J = 4.7 Hz, 1H), 3.93-3.46 (m, 12H), 3.40 (d, J = 11.8 Hz, 1H),
1.88 (d, J = 12.8 Hz, 1H), 1.79-1.68 (m, 1H), 1.60-1.45 (m, 1H),
1.31-1.16 (m, 1H) - two peaks are obscured on either side of DMSO
signal. 232 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.43 (s, 2H),
8.32 (s, 3H), 540 7.49 (d, J = 8.5 Hz, 2H), 7.38-7.07 (m, 5H), 6.88
(d, J = 8.7 Hz, 2H), 4.63-4.26 (m, 2H), 3.78 (m, 10H), 3.61 (m,
1H), 3.21-2.91 (m, 3H), 1.88-1.49 (m, 2H), 1.04 (d, J = 6.1 Hz,
1H). 234 4 .sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm 9.46 (br. s.,
1H), 8.38 (s, 540 2H), 8.20 (s, 1H), 7.53 (d, 2H, J = 8.0 Hz), 7.44
(d, 2H, J = 8.0 Hz), 7.33-7.29 (m, 2H), 7.22-7.18 (m, 1H), 6.91 (d,
2H, J = 8.0 Hz), 5.80 (s, 1H), 3.81 (br. s., 8H), 3.73 (t, 4H, J =
4.8 Hz), 3.04 (t, 4H, J = 4.8 Hz), 1.82 (s, 3H). 235 4 .sup.1H-NMR
(400 MHz, DMSO-d6) .delta. ppm 9.47 (br. s., 1H), 8.38 (s, 540 2H),
8.21 (s, 1H), 7.53 (d, 2H, J = 8.0 Hz), 7.45-7.43 (m, 2H),
7.33-7.29 (m, 2H), 7.22-7.19 (m, 1H), 6.91 (d, 2H, J = 8.0 Hz),
5.80 (s, 1H), 3.81 (br. s., 8H), 3.73 (t, 4H, J = 4.8 Hz), 3.04 (t,
4H, J = 4.8 Hz), 1.82 (s, 3H). 236 3 .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. ppm 8.53 (br. s., 1H), 8.26 (s, 1H), 8.19 (s,
2H), 7.53-7.51 (m, 2H), 7.41-7.40 (m, 2H), 7.30-7.29 (m, 2H),
7.24-7.15 (m, 3H), 3.86-3.73 (m, 11H), 3.39-3.30 (m, 3H), 3.07-3.05
(m, 1H), 2.81-2.79 (m, 1H), 2.32-2.22 (m, 1H), 1.96-1.75 (m, 3H).
237 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.67 (s, 1H), 8.31 (s,
2H), 541 8.25 (s, 1H), 7.55 (d, J = 15.1 Hz, 1H), 7.40 (s, 1H),
7.32-7.14 (m, 5H), 6.99 (t, J = 9.4 Hz, 1H), 3.88-3.71 (m, 17H),
2.96 (t, J = 4.7 Hz, 4H), 2.25 (s, 3H) - one peak is partially
obscured by water. 238 2 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.05 (s, 1H), 8.29 (s, 2H), 542 8.15 (s, 1H), 7.43-7.11 (m, 6H),
6.87 (dd, J = 12.5, 2.7 Hz, 1H), 6.75 (dd, J = 9.1, 2.7 Hz, 1H),
4.28-4.17 (m, 1H), 3.76 (m, 10H), 3.07 (d, J = 11.4 Hz, 1H), 2.75
(dt, J = 12.4, 4.0 Hz, 1H), 2.01 (s, 1H), 1.71-1.59 (m, 1H),
1.54-1.37 (m, 2H) - one peak obscured by DMSO signal. 239 2
.sup.1H-NMR (500 MHz, CDCl3) .delta. ppm 8.25 (s, 1H), 8.17 (s,
2H), 542 7.41 (d, 2H, J = 8.5 Hz), 7.13-7.10 (m, 2H), 7.00-6.93 (m,
4H), 3.96-3.86 (m, 9H), 3.78 (s, 2H), 3.28-3.24 (m, 1H), 3.08-3.03
(m, 3H), 2.48-2.27 (br, 1H), 1.95-1.91 (m, 1H), 1.84-1.76 (m, 2H),
1.72-1.60 (m, 2H). 240 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.43 (s, 1H), 8.31 (s, 2H), 542 8.20 (s, 1H), 7.50 (d, J = 8.4 Hz,
2H), 7.32-7.23 (m, 2H), 7.16-7.07 (m, 2H), 6.87 (d, J = 8.5 Hz,
2H), 4.78 (d, J = 4.7 Hz, 1H), 3.80 (m, 10H), 3.57 (m, 2H), 2.57
(td, J = 11.8, 3.2 Hz, 1H), 2.43 (dd, J = 11.3, 9.0 Hz, 1H), 1.87
(dt, J = 12.6, 4.3 Hz, 1H), 1.73 (ddd, J = 11.6, 7.9, 4.0 Hz, 1H),
1.60-1.45 (m, 1H), 1.29-1.17 (m, 1H). 241 3 This spectrum contains
some rotomers in the aromatic region: .sup.1H 542 NMR (400 MHz,
DMSO-d6) .delta. 9.62, 9.48 (s, 1H), 8.31, 8.27 (s, 2H), 8.15 (s,
1H), 7.82 (s, 1H), 7.48, 7.47 (s, 1H), 7.36-7.12 (m, 5H), 4.22-4.05
(m, 2H), 3.79 (m, 10H), 3.54-3.39 (m, 2H), 2.75-2.58 (m, 2H), 2.28
(q, J = 7.2 Hz, 2H), 2.02-1.89 (m, 1H), 1.72 (t, J = 10.3 Hz, 1H),
0.96 (t, J = 7.1 Hz, 3H). 242 3 .sup.1H-NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 9.00-8.74 (m, 1H), 8.31 (s, 2H), 8.14 (s, 1H),
7.58-7.41 (m, 1H), 7.30-7.27 (m, 2H), 7.24-7.17 (m, 3H), 4.83 (br.
s., 1H), 4.60-4.31 (m, 3H), 4.02-3.96 (m, 2H), 3.78 (s, 2H),
3.69-3.64 (m, 2H), 3.37-3.34 (m, 2H), 3.21-3.13 (m, 2H), 3.02-2.96
(m, 1H), 2.73-2.59 (m, 3H), 2.41-2.37 (m, 1H), 2.17, 2.12 (m, 3H),
1.15-1.01 (br, 3H). 243 1 .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 8.98, 8.73 (s, s, 1H), 8.31 (s, 2H), 8.14 (s, 1H),
7.53, 7.39 (s, s, 1H), 7.27 (br. s., 4H), 7.19-7.17 (m, 1H),
4.05-3.97 (m, 3H), 3.78-3.66 (m, 10H), 2.73-2.56 (m, 4H), 2.40-2.35
(m, 1H), 2.14-2.07 (m, 3H), 1.55 (d, 3H, J = 6.8 Hz). 244 4
.sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. ppm 8.33, 8.23 (br. s.,
s,
3H), 7.85, 7.76 (s, s, 1H), 7.58, 7.51 (s, s, 1H), 7.30-7.26 (m,
2H), 7.26-7.24 (m, 1H), 7.20-7.18 (m, 2H), 7.71, 6.60 (br. s., br.
s., 1H), 4.16-4.15 (m, 2H), 3.92-3.88 (m, 10H), 3.67-3.65 (m, 1H),
3.00-2.98 (m, 1H), 2.88-2.87 (m, 2H), 2.61-2.55 (m, 1H), 1.67 (s,
6H). 245 1 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.97,
8.71 (s, s, 1H), 8.13 (d, 1H, J = 6.5 Hz), 7.64 (s, 1H), 7.54, 7.40
(s, s, 1H), 7.28-7.17 (m, 5H), 6.31 (s, 2H), 4.01-3.98 (m, 2H),
3.76-3.65 (m, 13H), 2.74-2.60 (m, 3H), 2.41-2.36 (m, 1H), 2.16,
2.12 (m, 3H). 246 1 1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
9.62, 9.47 (s, s, 1H), 8.28, 8.18 (s, s, 1H), 8.02 (s, 1H), 7.83
(s, 1H), 7.50, 7.47 (s, s, 1H), 7.26 (t, 2H, J = 7.5 Hz), 7.19-7.15
(m, 3H), 4.13-4.01 (m, 2H), 3.87 (s, 3H), 3.87-3.65 (m, 12H),
3.42-3.36 (m, 1H), 2.72-2.57 (m, 3H), 2.37-2.33 (m, 1H). 247 1
.sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.66-9.52 (m, 1H),
8.32 (d, 2H, J = 7.5 Hz), 8.29, 8.16 (s, s, 1H), 8.02-7.83 (m, 1H),
7.54-7.48 (m, 1H), 7.30 (t, 2H, J = 7.5 Hz), 7.24 (d, 2H, J = 7.0
Hz), 7.20 (t, 1H, J = 7.5 Hz), 4.98-4.28 (m, 5H), 4.09-4.04 (m,
2H), 3.80 (s, 2H), 3.72-3.67 (m, 2H), 3.34-3.07 (m, 4H), 2.70-2.56
(m, 3H), 2.38-2.32 (m, 2H). 248 2 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.64-9.49 (m, 1H), 8.31 (d, 2H, J = 7.0
Hz), 8.28, 8.15 (s, s, 1H), 8.00, 7.83 (s, s, 1H), 7.53, 7.49, 7.47
(s, s, s, 1H), 7.30 (t, 2H, J = 7.5 Hz), 7.24 (d, 2H, J = 7.0 Hz),
7.19 (t, 1H, J = 7.5 Hz), 4.95-3.99 (m, 7H), 3.80 (s, 2H),
3.72-3.65 (m, 2H), 3.46-3.36 (m, 2H), 3.28-3.03 (m, 2H), 2.70-2.54
(m, 3H), 2.38-2.32 (m, 2H). 249 1 .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.64, 9.51 (s, s, 1H), 8.33 (s, 2H),
8.28, 8.16 (s, s, 1H), 7.83 (s, 1H), 7.49, 7.47 (s, s, 1H),
7.22-7.18 (m, 1H), 6.82-6.75 (m, 3H), 4.11-4.03 (m, 2H), 3.83-3.65
(m, 11H), 3.43-3.32 (m, 6H), 2.72-2.57 (m, 3H), 2.38-2.32 (m, 1H).
250 4 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.65, 9.52
(s, s, 1H), 8.38 (s, 2H), 8.28, 8.15 (s, s, 1H), 7.82 (s, 1H),
7.49-7.43 (m, 3H), 7.32-7.29 (m, 2H), 7.22-7.19 (m, 1H), 5.83 (s,
1H), 4.11-4.02 (m, 2H), 3.83-3.78 (m, 8H), 3.71-3.64 (m, 2H),
3.40-3.36 (m, 1H), 2.70-2.56 (m, 3H), 2.36-2.32 (m, 2H), 1.81 (s,
3H). 251 5 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
9.80-9.66 (m, 2H), 8.55 (s, 2H), 8.35 (s, 1H), 7.73, 7.72 (s, s,
2H), 7.35, 7.34 (s, s, 2H), 7.29-7.23 (m, 2H), 7.17-7.14 (m, 1H),
7.06-7.02 (m, 1H), 4.77-7.73 (m, 1H), 4.09 (dd, 1H, J = 12.0, 3.0
Hz), 3.97-3.90 (m, 9H), 3.35 (d, 1H, J = 11.5 Hz), 3.23 (d, 1H, J =
13.0 Hz), 3.12-2.95 (m, 2H). 252 3 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.78 (br. s., 1H), 8.33 (s, 2H), 8.29 (s,
1H), 7.73 (d, 2H, J = 8.5 Hz), 7.35 (d, 2H, J = 8.5 Hz), 7.30 (t,
2H, J = 7.5 Hz), 7.24 (d, 2H, J = 7.0 Hz), 7.20 (t, 1H, J = 7.0
Hz), 4.35 (s, 2H), 3.85-3.81 (m, 10H), 2.72 (s, 6H). 253 2 .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 9.64, 9.51 (s, 1H, rotamer), 548
8.50 (s, 2H), 8.17 (s, 1H), 7.83 (s, 1H), 7.49 (s, 1H), 7.24-6.82
(m, 4H), 4.93 (s, 2H), 4.18-3.97 (m, 2H), 3.96-3.55 (m, 11H),
2.78-2.57 (m, 4H), 2.43-2.21 (m, 1H). 254 4 .sup.1H-NMR (500 MHz,
CDCl.sub.3) .delta. ppm 8.82, 8.39 (s, s, 2H), 8.32, 8.22 (br. s.,
br. s., 1H), 7.83-7.74 (m, 1H), 7.61-7.59 (m, 1H), 7.52-7.45 (m,
5H), 7.30-7.27 (br, 1H), 4.14-4.13 (m, 2H), 3.95-3.82 (m, 11H),
3.62-3.58 (m, 1H), 2.94-2.81 (m, 3H), 2.58-2.54 (m, 1H). 255 5
.sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.68, 9.55 (s, s,
1H), 550 8.55 (s, 2H), 8.30, 8.18 (s, s, 1H), 7.83 (s, 1H), 7.50,
7.47 (s, s, 1H), 7.29-7.25 (m, 2H), 7.15 (td, 1H, J = 8.5, 2.0 Hz),
7.04 (t, J = 8.0 Hz, 1H), 4.14-4.03 (m, 2H), 3.94-3.82 (m, 8H),
3.72-3.65 (m, 2H), 3.39 (td, J = 11.0 Hz, J = 3.0 Hz 1H), 2.71-2.58
(m, 3H), 2.37-2.31 (m, 2H). 256 5 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.67, 9.53 (s, s, 1H), 550 8.54 (s, 2H),
8.30, 8.17 (s, s, 1H), 7.83 (s, 1H), 7.50, 7.47 (s, s, 1H),
7.28-7.25 (m, 2H), 7.17 (t, 2H, J = 8.0 Hz), 4.11-4.03 (m, 2H),
3.94-3.88 (m, 8H), 3.72-3.64 (m, 2H), 3.41-3.36 (m, 1H), 2.71-2.60
(m, 3H), 2.37-2.33 (m, 2H). 257 5 .sup.1H-NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 9.67, 9.54 (s, s, 1H), 8.56 (s, 2H),
8.30, 8.18 (s, s, 1H), 7.83 (s, 1H), 7.51, 7.47 (s, s, 1H),
7.38-7.33 (m, 1H), 7.04-6.96 (m, 3H), 4.14-4.03 (m, 2H), 3.95-3.90
(m, 8H), 3.72-3.65 (m, 2H), 3.42-3.37 (m, 1H), 2.71-2.60 (m, 3H),
2.37-2.31 (m, 2H). 258 5 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.66 (s, 1H), 8.40 (s, 1H), 551 8.18 (s, 1H), 7.82 (s, 1H),
7.57-7.45 (m, 3H), 7.31-7.20 (m, 2H), 4.07 (m, 2H), 3.90 (m, 8H),
3.75-3.63 (m, 3H), 3.47-3.24 (m, 3H), 2.74-2.54 (m, 3H), 2.40-2.28
(m, 1H). 259 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.65 (s, 1H),
8.30 (s, 2H), 551 8.25 (s, 1H), 8.20 (s, 1H), 7.82 (s, 2H), 7.46
(d, J = 8.1 Hz, 1H), 7.33-7.12 (m, 6H), 6.90 (d, J = 7.5 Hz, 1H),
3.87-3.75 (m, 16H), 2.75-2.63 (m, 3H), 2.65-2.54 (m, 1H), 2.45-2.35
(m, 2H), 2.29-2.20 (m, 2H), 2.08 (s, 6H), 1.89-1.81 (m, 1H),
1.66-1.55 (m, 1H). One peak is obscured by water peak and cannot be
accurately integrated 260 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.64 (s, 1H), 8.30 (s, 2H), 551 8.24 (s, 1H), 7.82 (s, 1H), 7.45
(s, 1H), 7.31-7.25 (m, 3H), 7.25-7.13 (m, 4H), 6.90 (d, J = 7.5 Hz,
1H), 3.92-3.67 (m, 10H), 3.57 (d, J = 13.1 Hz, 1H), 3.43 (d, J =
13.0 Hz, 1H), 2.74-2.54 (m, 3H), 2.44-2.32 (m, 1H), 2.28-2.16 (m,
1H), 2.05 (s, 6H), 1.90-1.78 (m, 1H), 1.66-1.52 (m, 1H). 261 3
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.64 (s, 1H), 8.30 (s, 2H),
551 8.24 (s, 1H), 7.81 (s, 1H), 7.45 (s, 1H), 7.32-7.23 (m, 3H),
7.26-7.13 (m, 4H), 6.90 (d, J = 7.6 Hz, 1H), 3.91-3.70 (m, 10H),
3.57 (d, J = 13.0 Hz, 1H), 3.44 (d, J = 13.0 Hz, 1H), 2.77-2.54 (m,
3H), 2.46-2.34 (m, 1H), 2.30-2.17 (m, 1H), 2.08 (s, 6H), 1.92-1.78
(m, 1H), 1.68-1.53 (m, 1H). 262 3 .sup.1H-NMR (500 MHz, CDCl.sub.3)
.delta. ppm 8.27 (s, 1H), 8.21 (s, 2H), 7.31-7.28 (m, 3H),
7.25-7.08 (m, 5H), 3.91-3.88 (m, 8H), 3.82-3.80 (m, 4H), 2.94-2.88
(m, 4H), 2.52 (s, 2H), 1.22 (s, 6H). 263 4 .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 9.43 (s, 1H), 8.25 (s, 2H), 552 8.19 (s, 1H), 7.49
(d, J = 8.3 Hz, 2H), 7.35-7.23 (m, 4H), 7.23-7.13 (m, 1H), 6.86 (d,
J = 8.6 Hz, 2H), 4.77 (s, 1H), 3.88-3.70 (m, 8H), 3.65-3.46 (m,
3H), 2.62-2.53 (m, 1H), 2.46-2.37 (m, 1H), 1.92-1.82 (m, 1H), 1.72
(dd, J = 10.4, 6.6 Hz, 1H), 1.63 (s, 6H), 1.52 (m, 1H), 1.24 (dt, J
= 13.2, 6.2 Hz, 2H). 265 1 .sup.1H NMR (400 MHz, DMSO-d6) .delta.
9.48 (s, 1H), 8.20 (s, 1H), 553 7.64 (s, 1H), 7.55 (d, J = 8.6 Hz,
2H), 7.31-7.11 (m, 5H), 6.89 (d, J = 8.5 Hz, 2H), 6.30 (s, 2H),
3.94 (dd, J = 9.6, 5.4 Hz, 1H), 3.85-3.58 (m, 10H), 2.97 (br.s,
1H), 2.37 (s, 3H), 2.21 (br.s, 1H), 1.96 (t, J = 10.3 Hz, 1H),
1.76-1.52 (m, 3H) one peak obscured by DMSO signal. 266 1 .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 9.48 (s, 1H), 8.20 (s, 1H), 553 7.64
(s, 1H), 7.56 (dd, J = 9.1, 3.0 Hz, 2H), 7.34-7.10 (m, 5H), 6.88
(d, J = 8.5 Hz, 2H), 6.30 (s, 2H), 3.93 (dd, J = 9.6, 5.3 Hz, 1H),
3.72 (dd, J = 41.7, 13.4 Hz, 10H), 2.96 (s, 1H), 2.36 (s, 3H), 2.19
(s, 1H), 1.95 (dq, J = 12.4, 8.2 Hz, 1H), 1.74-1.62 (m, 2H),
1.63-1.50 (m, 1H) one peak obscured by DMSO signal. 267 4
.sup.1H-NMR (400 MHz, DMSO-d6) .delta. ppm 9.65, 9.50 (s, s, 1H),
560 8.27 (s, 2H), 8.16 (s, 1H), 7.83 (s, 1H), 7.50, 7.47 (s, s,
1H), 7.30 (dd, 2H, J = 8.8, 6.0 Hz), 7.14-7.09 (m, 2H), 4.11-4.03
(m, 2H), 3.84-3.66 (m, 10H), 2.72-2.57 (m, 4H), 2.35-2.32 (m, 2H),
1.63 (s, 6H). 268 3 .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.58 (s,
1H), 8.31 (s, 2H), 562 8.25 (s, 1H), 7.42 (d, J = 6.3 Hz, 2H),
7.33-7.15 (m, 5H), 7.04 (d, J = 8.7 Hz, 1H), 3.89-3.71 (m, 12H),
2.90 (t, J = 5.9 Hz, 2H), 2.75 (t, J = 5.7 Hz, 2H) - one peak is
obscured by water signal. 269 4 .sup.1H-NMR (400 MHz, CDCl3)
.delta. ppm 8.36 (s, 2H), 8.20 (br. s., 1H), 562 7.75 (br. s., 1H),
7.57 (br. s., 1H), 7.42-7.38 (m, 2H), 7.05-7.00 (m, 2H), 4.15-4.11
(m, 2H), 3.90-3.84 (m, 11H), 3.62-3.56 (m, 1H), 2.93-2.79 (m, 2H),
2.58-2.53 (m, 1H), 1.93 (s, 3H). 270 4 .sup.1H-NMR (400 MHz,
DMSO-d6) .delta. ppm 9.64, 9.50 (s, s, 1H), 562 8.38 (s, 2H), 8.29,
8.16 (s, s, 1H), 7.83 (s, 1H), 7.49-7.44 (m, 3H), 7.14-7.10 (m,
2H), 5.89 (s, 1H), 4.11-4.02 (m, 2H), 3.84-3.66 (m, 10H), 2.71-2.57
(m, 3H), 2.38-2.32 (m, 1H), 1.81 (s, 3H). 271 1 .sup.1H-NMR (500
MHz, DMSO-d.sub.6) .delta. ppm 9.00, 8.75 (s, s, 1H), 8.30 (s, 2H),
8.25 (s, 1H), 8.15, 8.13 (s, s, 1H), 7.55, 7.41 (s, s, 1H),
7.28-7.25 (m, 2H), 7.13-7.10 (m, 2H), 4.07-3.98 (m, 3H), 3.78-3.70
(m, 13H), 3.41-3.37 (m, 1H), 2.80-2.62 (m, 3H), 2.44-2.42 (m, 1H),
2.16, 2.11 (s, s, 3H). 272 1 .sup.1H-NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 8.99, 8.75 (s, s, 1H), 8.29 (s, 2H), 8.20 (s, 1H),
8.15, 8.13 (s, s, 1H), 7.54, 7.40 (s, s, 1H), 7.33-7.25 (m, 2H),
7.17-7.12 (m, 2H), 4.03-3.96 (m, 2H), 3.81-3.69 (m, 12H), 3.45-3.33
(m, 1H), 2.78-2.62 (m, 3H), 2.45-2.36 (m, 2H), 2.15, 2.11 (s, s,
3H). 273 5 .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.03,
8.78 (s, s, 1H), 8.53 (s, 2H), 8.16-8.15 (m, 1H), 7.29, 7.24 (s, s,
1H), 7.27-7.24 (m, 2H), 7.19-7.15 (m, 2H), 4.02-3.96 (m, 2H),
3.86-3.64 (m, 10H), 3.37-3.35 (br. s., 2H), 2.74-2.56 (m, 3H),
2.41-2.37 (m, 1H), 2.17, 2.12 (s, s, 3H). 275 4 .sup.1H-NMR (400
MHz, CDCl3) .delta. ppm 8.35 (s, 2H), 8.25 (s, 1H), 571 7.43-7.35
(m, 4H), 7.00 (t, 2H, J = 8.8 Hz), 6.94 (d, 2H, J = 8.8 Hz),
7.00-6.93 (br., 1H), 3.96-3.93 (m, 1H), 3.92-3.80 (m, 8H),
3.28-3.24 (m, 1H), 3.09-3.03 (m, 3H), 2.35-2.15 (br., 1H),
1.95-1.91 (m, 1H), 1.83 (s, 3H), 1.82-1.70 (m, 3H). 276 4
.sup.1H-NMR (400 MHz, CDCl3) .delta. ppm 8.34 (s, 2H), 8.20 (br.
s., 1H), 576 7.71 (br. s., 0.5H), 7.41-7.38 (m, 2H), 7.04-6.99 (m,
2H), 6.50 (br. s., 0.5H), 4.09-4.08 (m, 2H), 3.87-3.85 (m, 10H),
3.62-3.56 (m, 1H), 2.98-2.83 (m, 3H), 2.65-2.59 (m, 1H), 2.22 (s,
3H), 1.92 (s, 3H). 277 4 .sup.1H-NMR (400 MHz, CDCl3) .delta. ppm
8.34 (s, 2H), 8.20 (br. s., 1H), 576 7.71 (br. s., 0.5H), 7.41-7.38
(m, 2H), 7.04-6.99 (m, 2H), 6.50 (br. s., 0.5H), 4.09-4.08 (m, 2H),
3.87-3.85 (m, 10H), 3.63-3.57 (m, 1H), 2.99-2.84 (m, 3H), 2.68-2.60
(m, 1H), 2.22 (s, 3H), 1.92 (s, 3H).
Biochemical Activity of Compounds
In order to assess the activity of chemical compounds against the
relevant kinase of interest, the Caliper LifeSciences
electrophoretic mobility shift technology platform is used.
Fluorescently labeled substrate peptide is incubated in the
presence of kinase and ATP so that a reflective proportion of the
peptide is phosphorylated. At the end of the reaction, the mix of
phosphorylated (product) and non-phosphorylated (substrate)
peptides are passed through the microfluidic system of the Caliper
EZ Reader 2, under an applied potential difference. The presence of
the phosphate group on the product peptide provides a difference in
mass and charge between those of the substrate peptide, resulting
in a separation of the substrate and product pools in the sample.
As the pools pass a LEDS within the instrument, these pools are
detected and resolved as separate peaks. The ratio between these
peaks therefore reflects the activity of the chemical matter at
that concentration in that well, under those conditions.
Kit wild type assay at Km: In each well of a 384-well plate, 0.2
ng/ul final (2 nM) of wild type Kit (Carna Bioscience 08-156) was
incubated in a total of 12.5 ul of buffer (100 mM HEPES pH 7.5,
0.015% Brij 35, 10 mM MgCl2, 1 mM DTT) with 1 uM Srctide
(5-FAM-GEEPLYWSFPAKKK-NH2) and 400 uM ATP at 25 C for 90 minutes in
the presence or absence of a dosed concentration series of compound
(1% DMSO final concentration). The reaction was stopped by the
addition of 70 ul of Stop buffer (100 mM HEPES pH 7.5, 0.015% Brij
35, 35 mM EDTA and 0.2% of Coating Reagent 3 (Caliper
Lifesciences)). The plate was then read on a Caliper EZReader 2
(protocol settings: -1.9 psi, upstream voltage -700, downstream
voltage -3000, post sample sip 35 s). Data was normalized to 0% and
100% inhibition controls and the IC50 or EC50 calculated using a
4-parameter fit using GraphPad Prism.
Kit D816V assay at Km: In each well of a 384-well plate, 0.04 ng/ul
(0.5 nM) of D816V Kit (Carna Bioscience 08-156) was incubated in a
total of 12.5 ul of buffer (100 mM HEPES pH 7.5, 0.015% Brij 35, 10
mM MgCl2, 1 mM DTT) with 1 uM Srctide (5-FAM-GEEPLYWSFPAKKK-NH2)
and 15 uM ATP at 25 C for 90 minutes in the presence or absence of
a dosed concentration series of compound (1% DMSO final
concentration). The reaction was stopped by the addition of 70 ul
of Stop buffer (100 mM HEPES pH 7.5, 0.015% Brij 35, 35 mM EDTA and
0.2% of Coating Reagent 3 (Caliper Lifesciences)). The plate was
then read on a Caliper EZReader 2 (protocol settings: -1.9 psi,
upstream voltage -700, downstream voltage -3000, post sample sip 35
s). Data was normalized to 0% and 100% inhibition controls and the
IC50 or EC50 calculated using a 4-parameter fit using GraphPad
Prism.
PDGFRA D842V assay at Km: In each well of a 384-well plate, 0.7
ng/ul (8 nM) of PDGFRA D842V (ProQinase 0761-0000-1) was incubated
in a total of 12.5 ul of buffer (100 mM HEPES pH 7.5, 0.015% Brij
35, 10 mM MgCl2, 1 mM DTT) with 1 uM CSKtide
(5-FAM-KKKKEEIYFFF-NH2) and 15 uM ATP at 25 C for 90 minutes in the
presence or absence of a dosed concentration series of compound (1%
DMSO final concentration). The reaction was stopped by the addition
of 70 ul of Stop buffer (100 mM HEPES pH 7.5, 0.015% Brij 35, 35 mM
EDTA and 0.2% of Coating Reagent 3 (Caliper Lifesciences)). The
plate was then read on a Caliper EZReader 2 (protocol settings:
-1.9 psi, upstream voltage -500, downstream voltage -3000, post
sample sip 38 s). Data was normalized to 0% and 100% inhibition
controls and the IC.sub.50 or EC.sub.50 calculated using a
4-parameter fit using GraphPad Prism.
Cellular Activity of Compound
HMC1.2 autophosphorylation assay:
10,000 HMC1.2 cells were incubated in 22 ul culture media
(phenol-red free IMDM, no serum) in each well of a 384-well plate
and serum starved overnight in a tissue culture incubator (5%
CO.sub.2, 37.degree. C.). A 10-point dose concentration series of
compound (25 uM-95.4 pM) were then added to the cells in a volume
of 3.1 ul to each well (0.25% DMSO final concentration). After 90
minutes, 6 ul of 5.times.AlphaLISA Lysis Buffer (Perkin Elmer)
supplemented with a protease and phosphatase inhibitor cocktail
(Cell Signaling Technologies) was added to each well and shaken at
450 rpm for 15 minutes at 4.degree. C. 10 ul of phospho-Y719 c-Kit
and total c-Kit antibodies (15 nM final concentration, Cell
Signaling Technologies) and 50 ug/ml AlphaLISA rabbit acceptor
beads (Perkin Elmer) were added to each well and shaken at 300 rpm
at room temperature for 2 hours. 10 ul of 100 ug/ml streptavidin
donor beads (Perkin Elmer) were added to each well, blocked from
light with aluminum adhesive and shaken at 300 rpm at room
temperature for 2 hours. Fluorescence signal was obtained on
Envision (Perkin Elmer) by AlphaScreen 384 well HTS protocol. Data
was normalized to 0% and 100% inhibition controls and the IC50 was
calculated using Four Parameter Logistic IC50 curve fitting.
The Table below shows the activity of compounds in a Mast cell
leukemia cell line, HMC 1.2. This cell line contains Kit mutated at
positions V560G and D816V resulting in constitutive activation of
the kinase. The following compounds were tested in an assay to
measure direct inhibition of Kit D816V kinase activity by assaying
Kit autophosphorylation at tyrosine 719 on the Kit protein.
The Table below shows the activity of compounds described herein,
against wild-type Kit, mutant Kit (the D816V mutant), and mutan
PDFGRA (the D852V mutant). In the Table below, for KIT D816V
activity and PDGFRA D842V, the following designations are used:
<1.00 nM=A; 1.01-10.0 nM=B; 10.01-100.0 nM=C; >100 nM=D; and
ND=not determined. For wild-type Kit activity, the following
designations are used: <10 nM=A; 11-100 nM=B; 100-1000 nM=C;
>1000 nM=D; and ND=not determined.
In the Table below, for cellular activity, the following
designations are used: <10 nM=A; 10.01-100 nM=B; 100.01-1000
nM=C; 1000-10000 nM=D, >10000.01 nM=E; and ND=not
determined.
TABLE-US-00003 Compound D816V WT PDFGRA Cellular Numer KIT KIT
D842V Activity 1 D ND 2 B ND D 3 D ND 4 A A A B 5 B C D 6 D ND E 7
B B D 8 C ND 9 C C 10 A B B C 11 C ND 12 A A A A 13 A A B 14 A B C
15 A B A B 16 A B C 17 B ND D 18 D ND E 19 C ND E 20 A ND B 21 B ND
C 22 B C C 23 B B C 24 B B C 25 A B A B 26 B B C 27 B ND B 28 A ND
B 29 A B 30 A B A B 31 A A B 32 B ND B 33 B ND B 34 C ND D 35 D ND
E 36 A A 37 D D 38 A B 39 A B 40 C B 41 A A B 42 B B B 43 A B B B
44 B B B B 45 B ND B 46 A A B B 47 A B A B 48 A A A B 49 A B B 50 B
B D 51 A B B 52 A A A B 53 A A A B 54 B C 55 A ND B 56 A A B 57 A B
B 58 B ND C 59 D C 60 A B B 61 A A A B 62 A ND B 63 D ND 64 A B B
65 A ND B 66 A ND B 67 C ND 68 B B A B 69 A A A A 70 B ND C 71 A ND
B 72 A ND B 73 A A A A 74 A B B 75 A A A B 76 A A B 77 B ND B 78 A
ND B 79 B B 80 B ND B 81 A ND B 82 A ND B 83 A A A B 84 A A B 85 A
B B 86 A B B 87 B ND B 88 A B A B 89 A B C 90 C ND 91 B ND D 92 A B
93 A B 94 B B B 95 A B D 96 A B C 97 A ND B 98 A A A B 99 B ND A
100 A ND B 101 A ND A A 102 A A A B 103 B B C 104 B B C 105 A B C
106 A B B C 107 A ND B 108 A ND B 109 A ND A 110 A ND B 111 B ND B
112 B ND C 113 B ND C 114 B C C 115 A A B 116 A A A B 117 A A A B
118 A A B 119 A A 120 A B 121 A B 122 A B 123 A A A A 124 A A A B
125 A ND B 126 B ND C 127 A A B 128 B B C 129 B ND B 130 B ND B 131
B ND B 132 A ND B 133 A ND B 134 A ND B 135 B ND B 136 B ND C 137 B
ND B 138 A ND B 139 B ND B 140 A 141 B ND B 142 C ND 143 B B C 144
B B C 145 C ND B 146 B ND B 147 B ND C 148 A A A B 149 A ND B B 150
B ND B 151 B ND B 152 A ND A 153 A ND B 154 B ND B 155 B ND B 156 A
A B 157 A A B C 158 B B C 159 B B C 160 B B C 161 A A B A 162 A A A
163 A A B 164 A A A 165 B C B B 166 A B B A 167 B C B 168 B B B 169
A B B 170 A B B 171 B ND B 172 A B 173 B ND B 174 A A A A 175 B ND
D 176 B ND B 177 B ND B 178 B ND C 179 A A A D 180 A B B 181 A A B
182 A B B 183 B ND B 184 B ND B 185 B ND B 186 A ND B 187 B C 188 B
B B 189 A ND B 190 B ND B 191 A A A 192 A ND A 193 A A 194 C ND 195
A A 196 A ND B 197 B ND C 198 A ND A 199 B ND B 200 B ND C 201 A B
202 B ND C 203 B ND B 204 B ND C 205 B ND B 206 B ND B 207 A B 208
A A 209 A ND B 210 A ND B 211 A ND B B 212 B B C 213 B ND B 214 A B
A B 215 A A B 216 A A A 217 A A A 218 A A B 219 A A B 220 A ND B
221 A ND A 222 B ND B 223 B B 224 A B 225 B B 226 B B B 227 B B 228
B ND C 229 A ND A 230 B ND D 231 A ND B 232 A ND B 233 B ND B 234 B
B 235 A A 236 A ND A 237 A B B 238 B ND C 239 B B B 240 B A 241 B
ND B 242 A ND B 243 B ND B 244 A ND B 245 A ND B
246 B ND D 247 C ND 248 A ND B 249 B ND B 250 A ND B 251 B ND C 252
B ND B 253 B ND B 254 B ND 255 A ND B 256 A ND B B 257 B ND C 258 B
ND B 259 A A A B 260 A A B 261 A A A B 262 A ND B 263 B B B 264 B
ND C 265 A A A 266 A A A 267 A B 268 B C C 269 A B 270 B 271 B ND B
A 272 A ND B 273 B ND B 274 C C 275 A B 276 B B 277 A B
Efficacy in an In Vivo Model
Compound 165 and Dasatinib were evaluated in a P815 mastocytoma
xenograft model. P815 tumor cells (ATCC, Manassas, Va., cat #
ATCC.RTM. TIB-64) were maintained in vitro as a suspension and
monolayer culture in RPMI1640 medium supplemented with 10% fetal
calf serum at 37.degree. C. in an atmosphere of 5% CO.sub.2 in air.
The tumor cells were sub-cultured twice weekly by trypsin-EDTA
treatment. The cells growing in an exponential growth phase were
harvested and counted for tumor inoculation.
Female BALB/c nude mice were used for the study. Each mouse was
inoculated subcutaneously in the right flank with the P815 tumor
cells (1.times.10.sup.6) in 0.1 ml of PBS for tumor development.
The treatments were started on day 6 after tumor inoculation when
the average tumor size reached approximately 89 mm.sup.3. The
testing article and vehicle were administrated to the mice
according to the regimen shown below.
TABLE-US-00004 Dose Dosing Volume Dosing Group n Treatment (mg/kg)
(ml/kg) Route Schedule* 1 13 Vehicle 0 10 p.o. BID .times. 10 2 10
Dasatinib 25 10 p.o. BID .times. 10 3 16 Compound 3 10 p.o. BID
.times. 10 165 4 16 Compound 10 10 p.o. BID .times. 10 165 5 16
Compound 30 10 p.o. BID .times. 10 165 6 16 Compound 100 10 p.o.
BID .times. 10 165 Note: *BID = twice per day.
Tumor sizes were measured every other day in two dimensions using a
caliper, and the volume was expressed in mm.sup.3 using the
formula: V=0.5 a.times.b.sup.2 where a and b were the long and
short diameters of the tumor, respectively. The tumor size was then
used for calculations of both T-C and T/C values. T-C was
calculated with T as the median time (in days) required for the
treatment group tumors to reach a predetermined size (e.g., 1000
mm.sup.3), and C as the median time (in days) for the control group
tumors to reach the same size. The T/C value (in percent) is an
indication of antitumor effectiveness; T and C are the mean volumes
of the treated and control groups, respectively, on a given
day.
TGI was calculated for each group using the formula: TGI
(%)=[1-(Ti-T0)/(Vi-V0)].times.100; Ti is the average tumor volume
of a treatment group on a given day, T0 is the average tumor volume
of the treatment group on the day of treatment start, Vi is the
average tumor volume of the vehicle control group on the same day
with Ti, and V0 is the average tumor volume of the vehicle group on
the day of treatment start. Tumor weight was measured at the
endpoint.
A statistical analysis of difference in tumor volume and tumor
weight among the groups was conducted on the data obtained at the
best therapeutic time point after the final dose (the 8.sup.th day
after the start of treatment). A one-way ANOVA was performed to
compare tumor volume and tumor weight among groups. All data were
analyzed using Prism 5.0. p<0.05 was considered to be
statistically significant.
Results. The tumor growth curves of different treatment groups are
shown in FIG. 1. Data points represent group mean tumor volume,
error bars represent standard error of the mean (SEM). As shown in
FIG. 1, Compound 165 was effective in inhibiting tumor growth.
Increasing the dose of Compound 165 enhanced the tumor inhibition
efficiency.
Thus, Compound 165, as a single agent, produced an observable
antitumor activity against the P815 mouse mastocytoma cancer
xenograft model in this study.
INCORPORATION BY REFERENCE
All publications and patents mentioned herein are hereby
incorporated by reference in their entirety as if each individual
publication or patent was specifically and individually indicated
to be incorporated by reference.
EQUIVALENTS
Those skilled in the art will recognize, or be able to ascertain
using no more than routine experimentation, many equivalents to the
specific embodiments of the invention described herein. Such
equivalents are intended to be encompassed by the following
claims.
* * * * *